E+H分析支架CYA251
GE Health care 柱子
GE HealthcareInstructions 71-5027-68 AH HisTrap affinity columns HisTrap HP, 1 ml and 5 mlHisTrap™ HP is a ready to use column, prepacked with precharged Ni Sepharose™ High Performance. This prepacked column is ideal for preparative purification of Histidine-tagged recombinant proteins by immobilized metal ion affinity chromatography (IMAC).The special design of the column, together with the high-performance matrix of the Ni Sepharose medium, provides fast, simple, and easy separations in a convenient format.Ni Sepharose High Performance has low nickel (Ni2+) ion leakage and is compatible with a wide range of additives used in protein purification. HP columns can be operated with a syringe, peristaltic pump, or liquid chromatography system such as ÄKTA™ design chromatography systems.CAUTION! Contains nickel. May produce an allergic reaction..1Pack size available by special order.Connector kit1Union 1/16” female/M6 male is also needed.2Union M6 female/1/16” male is also needed.Table of contents1.Description..............................................................................32.General considerations.....................................................63.Operation.................................................................................74.Optimization........................................................................115.Stripping and recharging..............................................126.Cleaning-in-place.............................................................137.Scaling-up............................................................................138.Storage..................................................................................149.Troubleshooting................................................................1410.Intended use.......................................................................1711.Ordering Information (18)Code No. Product No. supplied 17-5247-01 HisTrap HP 5 × 1 ml 17-5247-05 HisTrap HP 100 × 1 ml 117-5248-01 HisTrap HP 1 × 5 ml 17-5248-02 HisTrap HP 5 × 5 ml 17-5248-05HisTrap HP 100 × 5 ml 1Connectors supplied Usage No. supplied 1/16” male/luer femaleConnection of syringe to top of HiTrap column1Tubing connector flangeless/M6 femaleConnection of tubing (e.g. Peristaltic Pump P1) to bottom of HiTrap column 11Tubing connector flangeless/M6 male Connection of tubing (e.g. Peristaltic Pump P1) to top of HiTrap column 21Union 1/16” female/M6 male Connection to original FPLC™ System through bottom of HiTrap column1Union M6 female/1/16” maleConnection to original FPLC System through top of HiTrap column 1Stop plug female, 1/16”Sealing bottom of HiTrap column 2, 5 or 71DescriptionMedium propertiesHisTrap HP 1 ml and 5 ml columns are prepacked with NiSepharose High Performance, which consists of 34 µm highlycross-linked agarose beads with an immobilized chelating group.The medium has then been charged with Ni2+-ions.Several amino acids, for example histidine, form complexes with many metal ions. Ni Sepharose High Performance selectively binds proteins if suitable complex-forming amino acid residues areexposed on the protein surface.Additional histidines, such as in the case of (histidine)6 -tag,increase affinity for Ni2+ and generally make the histidine-tagged protein the strongest binder among other proteins in for example an E. coli extract.Column propertiesHisTrap HP columns are made of biocompatible polypropylenethat does not interact with biomolecules. Columns are deliveredwith a stopper on the inlet and a snap-off end on the outlet. Thecolumns have porous top and bottom frits that allow high flowrates. They cannot be opened or refilled.Columns can be operated with either a syringe and the supplied luer connector, a peristaltic pump, or a chromatography systemsuch as ÄKTA design.Note:To prevent leakage, ensure that the connector is tight.Table 1. HisTrap HP characteristics.1Dynamic binding capacity conditions:Note: Dynamic binding capacity is protein-dependent.2H 2O at room temperature 3Ni 2+-stripped mediumMatrixHighly cross-linked spherical agarose, 6%Average particle size 34 µmMetal ion capacity ~ 15 µmol Ni 2+/ml mediumDynamic binding capacity 1 At least 40 mg (histidine)6-tagged protein/ml medium Column volumes 1 ml or 5 ml Column dimensions i.d. × H:0.7 × 2.5 cm (1 ml) 1.6 × 2.5 cm (5 ml)Recommended flow rate 1 and 5 ml/min for 1 and 5 ml column respectivelyMax. flow rates 4 and 20 ml/min for 1 and 5 ml column respectively Max back pressure 20.3 MPa, 3 barCompatibility during use Stable in all commonly used buffers, reducingagents, denaturants, and detergents (s ee Table 2)Chemical stability 30.01 M HCl, 0.1 M NaOH. Tested for 1 week at 40°C.1 M NaOH, 70% acetic acid. Tested for 12 hours.2% SDS. Tested for 1 hour.30% 2-propanol. Tested for 30 min.Avoid in buffers Chelating agents, e.g. EDTA, EGTA, citrate (see Table 2)pH stability 3short term (at least 2 hours): 2 to 14 long term (≤ 1 week): 3 to 12Storage 20% ethanol Storage temperature4°C to 30°CSample:1 mg/ml (histidine)6-tagged pure protein (M r 28 000 or 43 000) inbinding buffer (Q B, 10% determination) or (histidine)6-tagged protein bound from E. coli extractColumn volume: 0.25 ml or 1 ml Flow rate: 0.25 ml/min or 1 ml/min Binding buffer: 20 mM sodium phosphate, 0.5 M NaCl, 5 mM imidazole, pH 7.4Elution buffer: 20 mM sodium phosphate, 0.5 M NaCl, 0.5 M imidazole, pH 7.4The Ni 2+-charged medium is compatible with all commonly used aqueous buffers, reducing agents, denaturants such as 6 M Gua-HCl and 8 M urea, and a range of other additives (see Table 2).Table 2. Ni Sepharose High Performance is compatible with the following compounds, at least at the concentrations given.1See Notes and blank run, p. 10–11.2Tested for 1 week at 40°C.3The strong chelator EDTA has been used successfully in some cases, at 1 mM.Generally, chelating agents should be used with caution (and only in the sample, not the buffers). Any metal-ion stripping may be counteracted by addition of a small excess of MgCl 2 before centrifugation/filtration of the sample. Note that stripping effects may vary with applied sample volume.Reducing agents 15 mM DTE 5 mM DTT20 mM ß-mercaptoethanol 5 mM TCEP10 mM reduced glutathione Denaturing agents 8 M urea 26 M guanidine hydrochloride 2Detergents2% Triton™ X-100 (nonionic)2% Tween™ 20 (nonionic)2% NP-40 (nonionic)2% cholate (anionic)1% CHAPS (zwitterionic)Other additives500 mM imidazole 20% ethanol 50% glycerol 100 mM Na 2SO 41.5 M NaCl 1 mM EDTA 360 mM citrate 3Buffer substances50 mM sodium phosphate, pH 7.4100 mM Tris-HCl, pH 7.4100 mM Tris-acetate, pH 7.4100 mM HEPES, pH 7.4100 mM MOPS, pH 7.4100 mM sodium acetate, pH 422General considerationsHisTrap HP is supplied precharged with Ni2+ ions. In general, Ni2+ is the preferred metal ion for purification of recombinant histidine-tagged proteins. Note, however, that in some cases it may be wise to test other metal ions, for example Zn2+ and Co2+, as the strength of binding depends on the nature of the histidine-tagged protein as well as the metal ion (see Optimization).We recommend binding at neutral to slightly alkaline pH (pH 7–8) in the presence of 0.5–1.0 M NaCl. Sodium phosphate buffers are often used. Tris-HCl can generally be used, but should be avoided in cases where the metal-protein affinity is very weak, since it may reduce binding strength. Avoid chelating agents such as EDTA or citrate in buffers, see Table2.Including salt, for example 0.5–1.0 M NaCl in the buffers andsamples, eliminates ion-exchange effects but can also have amarginal effect on the retention of proteins.Imidazole at low concentrations is commonly used in the binding and the wash buffers to minimize binding of host cell proteins. For the same reason, it is important to also include imidazole in thesample (generally, at the same concentration as in the washbuffer). At somewhat higher concentrations, imidazole may alsodecrease the binding of histidine-tagged proteins. The imidazole concentration must therefore be optimized to ensure the bestbalance of high purity (low binding of host cell proteins) and high yield (binding of histidine-tagged target protein). This optimalconcentration is different for different histidine-tagged proteins,and is usually slightly higher for Ni Sepharose High Performance than for similar IMAC media on the market (see Data File18-1174-40). Use highly pure imidazole; such imidazole givesessentially no absorbance at 280 nm.As alternatives to imidazole elution, histidine-tagged proteins can be eluted from the medium by several other methods orcombinations of methods – a lowering of pH within the range of2.5–7.5 can be used, for example. At pH values below 4, metal ionswill be stripped off the medium.Note:If the proteins are sensitive to low pH, we recommendcollection of the eluted fractions in tubes containing1 M Tris-HCl, pH 9.0 (60–200 µl/ml fraction) to restore thepH to neutral.Chelating agents such as EGTA or EDTA will also strip metal ionsfrom the medium and thereby cause protein elution, but the target protein pool will then contain Ni2+ ions. In this case, Ni2+ ions can be removed by desalting on a HiTrap™ Desalting, a PD-10 Desalting Column, or HiPrep™ 26/10 Desalting, (see Table3).Leakage of Ni2+ from Ni Sepharose High Performance is very low under all normal conditions, lower than for other IMAC mediatested. For applications where extremely low leakage duringpurification is critical, leakage can be even further reduced byperforming a blank run (see page 11). Likewise, a blank run should also be performed before applying buffers/ samples containingreducing agents (see page 11).Whatever conditions are chosen, HisTrap HP columns can beoperated with a syringe, peristaltic pump, or chromatographysystem.Note:If Peristaltic Pump P-1 is used, the maximum flow rate that can be run on a HisTrap HP 1 ml column is 3 ml/min.3OperationBuffer preparationWater and chemicals used for buffer preparation should be of high purity. Filter buffers through a 0.22 µm or a 0.45 µm filter beforeuse.Use high purity imidazole as this will give very low or noabsorbance at 280 nm.If the recombinant histidine-tagged protein is expressed asinclusion bodies, include 6 M Gua-HCl or 8 M urea in all buffers and sample. On-column refolding of the denatured protein may bepossible.Recommended conditionsSample preparationFor optimal growth, induction, and cell lysis conditions for your recombinant histidine-tagged clones, please refer to established protocols.Adjust the sample to the composition and pH of the binding buffer by: Adding buffer, NaCl, imidazole, and additives fromconcentrated stock solutions; by diluting the sample with binding buffer; or by buffer exchange, (see Table 3). Do not use strong bases or acids for pH-adjustment (precipitation risk). Filter the sample through a 0.22 µm or a 0.45 µm filter and/or centrifuge it immediately before applying it to the column.To prevent the binding of host cell proteins with exposed histidine, it is essential to include imidazole at a low concentration in the sample and binding buffer (see Optimization).Binding buffer: 20 mM sodium phosphate, 0.5 M NaCl,20–40 mM imidazole, pH 7.4 (The optimal imidazole concentration is protein-dependent; 20–40 mM is suitable for many proteins.)Elution buffer:20 mM sodium phosphate, 0.5 M NaCl, 500 mM imidazole, pH 7.4 (The imidazole concentration required for elution is protein-dependent).Table 3. Prepacked columns for desalting and buffer exchangePurification1Fill the syringe or pump tubing with distilled water. Remove the stopper and connect the column to the syringe (use the luerconnector provided), laboratory pump or chromatographysystem tubing “drop-to-drop” to avoid introducing air into thesystem.2Remove the snap-off end at the column outlet.3Wash the column with 3–5 column volumes of distilled water.4Equilibrate the column with at least 5 column volumes of binding buffer. Recommended flow rates are 1 ml/min or5 ml/min for the 1 and 5 ml columns respectively.In some cases a blank run is recommended before finalequilibration/ sample application (see page 11).5Apply the pretreated sample using a syringe or a pump.6Wash with binding buffer until the absorbance reaches a steady baseline (generally, at least 10–15 column volumes).Note:Purification results are improved by using imidazole insample and binding buffer (see Optimization).7Elute with elution buffer using a one-step or linear gradient.Five column volumes are usually sufficient if the protein ofinterest is eluted by a one-step gradient. A shallow gradient, forexample a linear gradient over 20 column volumes or more,may separate proteins with similar binding strengths.Note:If imidazole needs to be removed from the protein, use HiTrap Desalting, a PD-10 Desalting Column, or HiPrep26/10 Desalting depending on the sample volume (seeTable3).Note:Ni Sepharose High Performance is compatible withreducing agents. However, removal of any weakly boundNi2+ ions by performing a blank run without reducingagents (as described on page 11) before applying buffer/sample including reducing agents is recommended. Do notleave HisTrap HP columns with buffers including reducingagents when not in use.Note:Leakage of Ni2+ from Ni Sepharose High Performance is low under all normal conditions. The leakage is lower than forother IMAC media tested (see Data File Ni Sepharose HighPerformance, 18-1174-40). For very critical applications,leakage during purification can be even further diminishedby performing a blank run (as described below) beforeloading sample.Blank run:Use binding buffer and elution buffer without reducing agents.1Wash the column with 5 column volumes of distilled water (to remove the 20% ethanol).2Wash with 5 column volumes of elution buffer.3Equilibrate with 10 column volumes of binding buffer.4OptimizationImidazole at low concentrations is commonly used in the binding and the wash buffers to minimize binding of host cell proteins. For the same reason, it is important to also include imidazole in thesample (generally, at the same concentration as in the washbuffer). At somewhat higher concentrations, imidazole may alsodecrease the binding of histidine-tagged proteins. The imidazole concentration must therefore be optimized to ensure the bestbalance of high purity (low binding of host cell proteins), and high yield (binding of histidine-tagged target protein). This optimalconcentration is different for different histidine-tagged proteins,and is usually slightly higher for Ni Sepharose High Performance than for similar IMAC media on the market (see Data File NiSepharose High Performance, 18-1174-40). Finding the optimalimidazole concentration for a specific histidine-tagged protein is a trial-and-error effort, but 20–40 mM in the binding and washbuffers is a good starting point for many proteins. Use a high purity imidazole, such imidazole gives essentially no absorbance at280 nm.Ni2+ is usually the first choice metal ion for purifying most(histidine)6-tagged recombinant proteins from nontagged host cell proteins, and also the ion most generally used. Nevertheless, it isnot always possible to predict which metal ion will be best for agiven protein. The strength of binding between a protein and ametal ion is affected by several factors, including the length,position, and exposure of the affinity tag on the protein, the type of ion used, and the pH of buffers, so some proteins may be easier to purify with ions other than Ni2+.A quick and efficient way to test this possibility and optimizeseparation conditions is to use HiTrap IMAC HP 1 ml columns,which are packed with IMAC Sepharose High Performance (notcharged with metal ions). Each column can be charged withdifferent metal ions, for example Cu2+, Co2+, Zn2+, Ca2+, or Fe2+.Instructions are included with each column.A study to compare the purification of six (histidine)6-taggedrecombinant proteins, including three variants of maltose-binding protein, with different metal ions has indicated that Ni2+ generally gives best selectivity between (histidine)-tagged and nontaggedhost-cell proteins (see Application Note 18-1145-18).5Stripping and rechargingNote:The column does not have to be stripped and recharged between each purification if the same protein is going to bepurified; it is sufficient to strip and recharge it after 5–7purifications, depending on the cell extract, extract volume,target protein, etc.Recommended stripping buffer: 20 mM sodium phosphate,0.5 M NaCl, 50 mM EDTA, pH 7.4Strip the column by washing with at least 5–10 column volumes of stripping buffer. Wash with at least 5–10 column volumes ofbinding buffer and 5–10 column volumes of distilled water before recharging the column.Recharge the water-washed column by loading 0.5 ml or 2.5 ml of0.1 M NiSO4 in distilled water on HisTrap HP 1 ml and 5 ml column,respectively. Salts of other metals, chlorides, or sulfates, may also be used (see“Optimization”). Wash with 5 column volumes distilled water, and 5 column volumes binding buffer (to adjust pH) before storage in 20% ethanol.6Cleaning-in-placeWhen an increase in back-pressure is seen, the column can becleaned. Before cleaning, strip off Ni2+ ions using therecommended procedure described above.After cleaning, store in 20% ethanol (wash with 5 column volumes) or recharge with Ni2+ prior to storage in ethanol.The Ni2+-stripped column can be cleaned by the followingmethods;•Remove ionically bound proteins by washing the column with several column volumes of 1.5 M NaCl; then wash the columnwith approximately 10 column volumes of distilled water.•Remove precipitated proteins, hydrophobically bound proteins, and lipoproteins by washing the column with 1 MNaOH, contact time usually 1–2 hours (12 hours or more forendotoxin removal). Then wash the column withapproximately 10 column volumes of binding buffer, followedby 5–10 column volumes of distilled water.•Remove hydrophobically bound proteins, lipoproteins, and lipids by washing the column with 5–10 column volumes 30%isopropanol for about 15–20 minutes. Then wash the columnwith approximately 10 column volumes of distilled water.Alternatively, wash the column with 2 column volumes ofdetergent in a basic or acidic solution. Use, for example,0.1–0.5% nonionic detergent in 0.1 M acetic acid, contact time1–2 hours. After treatment, always remove residual detergentby washing with at least 5 column volumes of 70% ethanol.Then wash the column with approximately 10 column volumesof distilled water.7Scaling-upTwo or three HisTrap HP 1 ml or 5 ml columns can be connected in series for quick scale-up (note that back-pressure will increase).Ni Sepharose High Performance, the medium prepacked inHisTrap HP columns, is supplied preswollen in 25 and 100 ml labpacks (see Ordering Information). An alternative scale-up strategy is thus to pack the medium in empty columns – Tricorn™ and XK columns are suitable for this purpose.8StorageStore HisTrap HP columns in 20% ethanol at 4°C to 30°C.9TroubleshootingThe following tips may be of assistance. If you have any furtherquestions about your HisTrap HP column, please visit/hitrap, contact our technical support, or your local representative.Note:When using high concentrations of urea or Gua-HCl,protein unfolding generally takes place. Refolding on-column (or after elution) is protein-dependent.Tips:To minimize dilution of the sample, solid urea or Gua-HCl can be added.Tips:Samples containing urea can be analyzed directly by SDS-PAGE whereas samples containing Gua-HCl must be buffer-exchangedto a buffer with urea before SDS-PAGE.Column has clogged:•Cell debris in the sample may clog the column. Clean the column according to the section Cleaning-in-place.•It is important to centrifuge and/or filter the sample through a0.22 µm or a 0.45 µm filter, see Sample preparation. Sample is too viscous:•If the lysate is very viscous due to a high concentration of host nucleic acid, continue sonication until the viscosity is reduced,and/or add DNase I to 5 µg/ml, Mg2+ to 1 mM, and incubate onice for 10–15 min. Alternatively, draw the lysate through asyringe needle several times.Protein is difficult to dissolve or precipitates during purification:•The following additives may be used: 2% Triton X-100, 2% Tween 20, 2% NP-40, 2% cholate, 1% CHAPS, 1.5 M NaCl,50% glycerol, 20 mM ß-mercaptoethanol, 1–3 mM DTT or DTE(up to 5 mM is possible but depends on the sample and thesample volume), 5 mM TCEP, 10 mM reduced glutathione, 8 Murea, or 6 M Gua-HCl. Mix gently for 30 min to aid solubilizationof the tagged protein (inclusion bodies may require longermixing). Note that Triton X-100 and NP-40 (but not Tween)have a high absorbance at 280 nm. Furthermore, detergentscannot be easily removed by buffer exchange.No histidine-tagged protein in the purified fractions:•Elution conditions are too mild (histidine-tagged protein still bound: Elute with an increasing imidazole gradient ordecreasing pH to determine the optimal elution conditions.•Protein has precipitated in the column: For the nextexperiment, decrease amount of sample, or decrease proteinconcentration by eluting with linear imidazole gradient insteadof imidazole steps. Try detergents or changed NaClconcentration, or elute under denaturing (unfolding)conditions (add 4–8 M urea or 4–6 M Gua-HCl).•Nonspecific hydrophobic or other interaction: Add a nonionic detergent to the elution buffer (e.g. 0.2% Triton X-100)or increase the NaCl concentration.•Concentration of imidazole in the sample and/or binding buffer is too high: The protein is found in the flow-throughmaterial. Decrease the imidazole concentration.•Histidine-tag may be insufficiently exposed: The protein is found in the flowthrough material; perform purification ofunfolded protein in urea or Gua-HCl as for inclusion bodies.•Buffer/sample composition is incorrect: The protein is found in the flowthrough material. Check pH and composition ofsample and binding buffer. Ensure that chelating or strongreducing agents are not present in the sample at too highconcentration, and that the concentration of imidazole is nottoo high.SDS-PAGE of samples collected during the preparation of thebacterial lysate may indicate that most of histidine-tagged protein is located in the centrifugation pellet. Possible causes andsolutions are:•Sonication may be insufficient: Cell disruption may be checked by microscopic examination or monitored bymeasuring the release of nucleic acids at A260. Addition oflysozyme (up to 0.1 volume of a 10 mg/ml lysozyme solution in25 mM Tris-HCl, pH 8.0) prior to sonication may improveresults. Avoid frothing and overheating as this may denaturethe target protein. Over-sonication can also lead tocopurification of host proteins with the target protein.•The protein may be insoluble (inclusion bodies): The protein can usually be solubilized (and unfolded) from inclusion bodiesusing common denaturants such as 4–6 M Gua-HCl, 4–8 Murea, or strong detergents. Prepare buffers containing 20 mMsodium phosphate, 8 M urea, or 6 M Gua-HCl, and suitableimidazole concentrations, pH 7.4–7.6. Buffers with urea shouldalso include 500 mM NaCl. Use these buffers for samplepreparation, as binding buffer and as elution buffer. Forsample preparation and binding buffer, use 10–20 mMimidazole or the concentration selected during optimizationtrials (including urea or Gua-HCl).The protein is collected but is not pure (multiple bands on SDS polyacrylamide gel):•Partial degradation of tagged protein by proteases: Add protease inhibitors (use EDTA with caution, see Table2).•Contaminants have high affinity for nickel ions: Elute with a stepwise or linear imidazole gradient to determine optimalimidazole concentrations to use for binding and for wash; addimidazole to the sample in the same concentration as thebinding buffer. Wash before elution with binding buffercontaining as high concentration of imidazole as possible,without causing elution of the tagged protein. A shallowimidazole gradient (20 column volumes or more), mayseparate proteins with similar binding strengths. If optimizedconditions do not remove contaminants, further purificationby ion exchange chromotography (HiTrap Q HP orHiTrap SP HP) and/or gel filtration (Superdex™ Peptide,Superdex 75 or Superdex 200) may be necessary.•Contaminants are associated with tagged proteins: Add detergent and/or reducing agents before sonicating cells.Increase detergent levels (e.g. up to 2% Triton X-100 or2% Tween 20), or add glycerol (up to 50%) to the wash bufferto disrupt nonspecific interactions.Histidine-tagged protein is eluted during sample loading/wash:•Buffer/sample composition is incorrect: Check pH and composition of sample and binding buffer. Ensure thatchelating or strong reducing agents are not present in thesample at a too high concentration, and that theconcentration of imidazole is not too high.•Histidine-tag is partially obstructed: Purify under denaturing conditions (use 4–8 M urea or 4–6 M Gua-HCl).•Column capacity is exceeded: Join two or three HisTrap HP1 ml columns together or change to a HisTrap HP 5 ml column. 10Intended useThe HisTrap HP is intended for research use only, and shall not be used in any clinical or in vitro procedures for diagnostic purposes.11Ordering InformationProduct No. supplied Code No.HisTrap HP 5 × 1 ml 17-5247-01100 × 1 ml1 17-5247-051 × 5 ml 17-5248-015 × 5 ml 17-5248-02100 × 5 ml1 17-5248-05 Related products No. supplied Code No.100 ml 17-5268-02 HiTrap Desalting 5 × 5 ml 17-1408-01100 × 5 ml1 11-0003-29 PD-10 Desalting Column 30 17-0851-01 HiPrep 26/10 Desalting 1 × 53 ml 17-5087-014 × 53 ml 17-5087-02HisTrap FF 5 × 1 ml 17-5319-01100 × 1 ml1 17-5319-025 × 5 ml 17-5255-01100 × 5 ml1 17-5255-02 HisTrap FF crude 5 × 1 ml 11-0004-58100 × 1 ml1 11-0004-595 × 5 ml 17-5286-01100 × 5 ml1 17-5286-02 HisTrap FF crude Kit 1 kit 28-4014-77 HisPrep™ FF 16/10 1 × 20 ml 28-9365-511Pack size available by special order.1 One connector included in each HiTrap package.2 Two, five, or seven stop plugs female included in HiTrap packages depending on the product.3One fingertight stop plug is connected to the top of each HiTrap column at delivery.AccessoriesNo. Supplied Code No.1Tubing connector flangeless/M6 female 12 18-1003-68Tubing connector flangeless/M6 male 1 2 18-1017-98Union 1/16” female/M6 male 1 6 18-1112-57Union M6 female /1/16” male 1 5 18-3858-01Union luerlock female/M6 female 218-1027-12HiTrap/HiPrep, 1/16” male connector for ÄKTA design8 28-4010-81Stop plug female, 1/16”2 5 11-0004-64Fingertight stop plug, 1/16”35 11-0003-55Related literatureCode No.Recombinant Protein Purification Handbook, Principles and Methods18-1142-75Affinity Chromatography Handbook, Principles and Methods18-1022-29Affinity Chromatography, Columns and Media Selection Guide18-1121-86Ni Sepharose and IMAC Sepharose, Selection Guide 28-4070-92HiTrap Column Guide18-1129-81。
hilic色谱柱 寡糖
hilic色谱柱寡糖
HILIC色谱柱是一种亲水性交换色谱柱，用于分离和分析亲水性分子，例如糖类、寡糖。
寡糖是由2-9个单糖单元组成的糖分子，具有重要的生物学功能和应用价值。
在糖类的分析中，HILIC色谱柱常被用于寡糖的分离和定量分析。
HILIC色谱使用极性固定相和较强的有机溶剂作为流动相，通过样品分子与固定相之间的亲水性相互作用来实现分离。
在HILIC色谱柱上，寡糖在流动相中与固定相发生相互作用，不同寡糖之间的亲水性差异导致它们在柱上有不同的保留时间，从而实现分离。
使用HILIC色谱柱进行寡糖分析时，常常需要选择合适的流动相和柱温来优化分离性能。
同时，可以通过检测器测量吸光度、折射率或电导率等来检测和定量寡糖。
总之，HILIC色谱柱是一种常用的分离和分析寡糖的工具，可以提供高分辨率和灵敏度的分析结果。
VB5101 阀支架安装及使用说明书
VB5101阀支架安装及使用说明书大连依利特分析仪器有限公司生产的VB5101阀支架设计新颖、方便实用，不仅可以用于安装进样阀，还可以作为高效液相色谱柱的支架。
VB5101阀支架适合于安装Rheodyne公司7725i进样阀，可以十分方便地将支架安装于大连依利特分析仪器有限公司生产的iChrom系列产品中泵或色谱柱恒温箱的右侧。
一、结构
VB5101阀支架的结构示意图如图1所示。
二、安装方法
当收到VB5101阀支架后，对照图1中明细验收所购的物品，所有的零配件应完整，所有的物品表面应无明显的碰撞伤痕。
如果存在任何问题请与当地的代理商或本公司直接联系。
与仪器的连接：按照图1组装支架后，用4个圆柱头内六角螺钉M4×10把2角钢拧紧在仪器框架底部的四个螺纹孔上，安装完成后的效果如图2所示。
进样阀的安装：用进样阀自带的内六角扳手将进样阀的手柄卸下，根据实际使用需要，选择3阀支架上合适的孔位，用进样阀随机所带的螺钉将阀固定在3阀支架上即可。
色谱柱的固定：旋松9色谱柱固定手扭，将色谱柱放于1滑块凹槽内，旋紧9色谱柱固定手扭即可。
为了减少进样后的样品扩散，色谱柱应以向上的方向使用。
图1.VB5101
阀支架结构示意图
图2.VB5101阀支架安装于仪器右侧示意图大连依利特分析仪器有限公司
公司地址：大连市高新园区七贤岭学子街2-2号
公司电话：*************（总机）；客户服务专线：4006635483（Elite）
公司传真：*************
公司邮箱：******************
公司网址：
DEAIC＠
2017.06.15。
e+H浊度仪样本
Turbimax CUS51D
浊度和悬浮固体浓度传感器 浸入式安装，用于低、中、高浊度和悬浮固体浓度测量
应用 Turbimax CUS51D 传感器适用于各种污水处理场合。 • 出水口浊度测量 • 活性污泥池和回流污泥中的悬浮固体浓度测量 • 污泥处理过程中的悬浮固体浓度测量 • 出水口的过滤性固体浓度测量 优势 • 传感器基于多种测量原理工作 (90° 散射光测量原理、
传感器的工厂标定为 “ 福马肼 ” 标定，并由此确定过滤性固体的 “ 高岭土 ” 标定。 其他类型的预先标定将根据相应介质进行优化。 多达五点标定。
应用
出厂福马肼标定 出厂高岭土标定 二氧化硅 ( SiO2) 模式 二氧化钛 ( TiO2) 模式 活性污泥模式 回流污泥模式 消化污泥 / 淤泥模式
推荐的标定范围
通过污水测量安装支架安装
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安装角度为 90°。
通过 CYA251 流通式安装支架安装
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Endress+Hauser
Turbimax CUS51D 管道安装
环境温度 储存温度 防护等级 过程温度 过程压力 最小流量
下图介绍了传感器在管道上的各种安装位置，并标识该安装位置是否合理。
135° 散射光测量原理和四脉冲光束测量原理 )，可以根 据实际测量任务择优选择 • 传感器已进行工厂标定 ( 基本福马肼 )。已经针对用户 的具体订购信息 ( 例如：活性污泥 ) 进行出厂预标定， 调试快速、简便 • 采用标准信号传输 (Memosens 技术 ) 方式，传感器 “ 即 插即用 ” • 智能型传感器 - 传感器中储存所有特征参数和标定值 • 用户可以自行进行传感器标定，有多达五点的安全可 靠实验室标定或现场标定
Endress+Hauser Memosens Wave CAS80E 全光谱传感器操作手册说明书
Products Solutions Services操作手册Memosens Wave CAS80E 全光谱传感器数字式全光谱水质分析传感器BA02005C/28/ZH/05.22-00715972932022-04-25Memosens Wave CAS80E 全光谱传感器目录Endress+Hauser 3目录1文档信息 (4)1.1安全图标 (4)1.2信息图标 (4)2基本安全指南 (5)2.1人员要求 (5)2.2指定用途 (5)2.3工作场所安全 (5)2.4操作安全 (5)2.5产品安全 (6)3产品描述 (7)3.1产品设计 (7)4到货验收和产品标识 (9)4.1到货验收 (9)4.2产品标识 (9)4.3供货清单 (10)5安装 (11)5.1安装要求 (11)5.2安装设备 (13)5.3安装后检查 (19)6电气连接 (20)6.1连接设备 (20)6.2确保防护等级 (21)6.3连接后检查 (21)7调试 (23)7.1功能检查 (23)8操作 (24)8.1基于过程条件调节测量设备 (24)8.2循环清洗 (27)9诊断和故障排除 (28)9.1常见故障排除 (28)10维护 (29)10.1维护计划 (29)10.2维护任务 (29)11维修 (31)11.1概述 (31)11.2备件 (31)11.3返厂 (31)11.4处置 (31)12附件..............................3212.1设备专用附件........................3213技术参数..........................3413.1输入...............................3413.2电源...............................3513.3性能参数............................3513.4环境条件............................3913.5过程条件............................3913.6机械结构............................39索引.. (41)文档信息Memosens Wave CAS80E 全光谱传感器4Endress+Hauser1 文档信息1.1 安全图标1.2 信息图标附加信息，提示允许或推荐的操作禁止或不推荐的操作参见设备文档参考页面参考图 操作结果1.2.1 设备上的图标参见设备文档资料此类产品不可作为未分类城市垃圾废弃处置。
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Flowfit CYA251
流通式安装支架 用于安装硝酸盐 /SAC、浊度和溶解氧传感器
应用 CYA251 流通式安装支架设计用于安装直径为 40 mm (1.57") 的传感器。适用于管道传输过程介质或采 样的所有应用，用于密闭管道。 主要应用场合如下： • 饮用水 • 地表水监测 • 分析站 • 过程应用 • 污水处理
• SAC 或硝酸盐测量 CAS51D • 溶解氧测量 COS31、 COS41、 COS51D、 COS 61 和 COS61D • 浊度测量 CUS31、 CUS41 和 CUS51D
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Flowfit CYA251
订购信息
产品选型表 通过下列方式获取产品的详细订购信息： • 使用 Endress+Hauser 公司网页上的产品选型软件： 选择国家 产品 选择仪表 功能页面：产品选型 • 咨询 Endress+Hauser 当地销售中心：/worldwide 产品选型软件：产品选型工具 • 最新设置参数 • 取决于设备类型：直接输入测量点参数，例如：测量范围或显示语言 • 自动校验排他选项 • 自动生成订货号及其明细， PDF 文件或 Excel 文件输出 • 通过 Endress+Hauser 在线商城直接订购 供货清单 供货清单包括： • 流通式安装支架 ( 包含传感器适配接口和过程连接 ) • 订购的附件 • 带止回阀的适配接头 ( 仅适用于清洗功能，压缩空气采用 6 mm 连接 ) • 润滑脂， 1 管 ( 适用于 O 型圈 ) • 《操作手册》 出厂时，安装支架上的清洗口和清洗连接均使用插塞密封。
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Flowfit CYA251
清洗
流通式安装支架可以选配清洗喷头。在 G¼ 连接处接入压缩空气或水，可以实现清洗功能。 G¼ 清洗连接可以连接： • 压缩空气，通过密闭止回阀 (G¼ ...6 mm)，或 •水
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可选清洗功能 1 2 3 清洗连接 清洗喷头，带 G¼ 螺纹连接 插塞
优势 • • • • • • 便捷的壁式安装或柱式安装 安装灵活 正确安装后可以实现自排气 内部容积小，响应时间短 通过清洗喷头实现自动清洗 ( 压缩空气或水 ) 通过清洗端口进行手动清洗 ( 取决于传感器 )
TI00495C/28/zh/02.12
Flowfit CYA251
功能与系统设计
测量原理 完整的测量系统器，例如：CAS51D • 变送器，例如：Liquiline CM442 • 测量电缆
连接浊度和溶解氧传感器时，可以通过下部清端口进行手动清洗。但是，连接 CAS51D 传感器， 无法进行此类清洗。
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安装条件
安装指南 进行旁通管流量测量时，压力 p1 必须高于 p2。因此，必须在主管道中安装节流孔板。
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连接示意图：在包含旁通管和孔板的主管道中安装
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CYA251，带传感器适配接头 1 传感器适配接头
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Flowfit CYA251
壁式安装支架
CYA251 的壁式安装套件 • 订货号：71144369
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壁式安装套件 1 悬挂螺栓 STST 10 x 60 mm
Endress+Hauser
COS61D COS61 COS31 326 (12.83)
CUS31 CUS41 325 (12.80)
CUS51D
Z mm(inch)
332 (13.07)
过程连接 X mm(inch) Y mm(inch)
NPT ¾" 83 (3.27) 50.5 (1.99)
Rp ¾ 83 (3.27) 50.5 (1.99)
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测量系统的结构示意图
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过程管道 Liquiline CM442 变送器 测量电缆 回水管道，带截止阀 CYA251 流通式安装支架，已安装 40 mm 传感器 进水管道，带截止阀
部件
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3
1
4
5
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流通式安装支架的部件示意图 1 2 3 4 5 进水口过程连接 传感器适配接头 ( 与传感器类型相关 ) 清洗连接 ( 清洗喷头可选 ) 出水口过程连接 清洗口
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壁式安装支架 1 悬挂螺栓 STST 10 x 60 ( 标准供货件 )
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环境条件
环境温度 储存温度 0...50 °C (32...120 °F) 0...50 °C (32...120 °F)
过程条件
过程温度 过程压力 0...50 °C (32...120 °F)，不结冰 20 °C (68 °F) 时， max. 6 bar (87 psi) 50 °C (122 °F) 时， max. 4 bar (58 psi)
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Flowfit CYA251
附件
清洗喷头 CYA251 的清洗喷头 • 用于自动清洗传感器 • 订货号：71144328 ( 适用于 CAS51D 传感器 (2 mm 或 8 mm 光程 )) • 订货号：71144330 ( 适用于 CAS51D 传感器 (40 mm 光程 )) • 订货号：71144331 ( 适用于 CUS51D、 COS51D、 COS61D、 COS61、 COS31、 COS41 传感器 ) • 订货号：71144332 ( 适用于 CUS31、 CUS41 传感器 )
DN 25 83 (3.27) 50.5 (1.99)
ANSI 1" 50 (1.97) 115 (4.53)
D20 软管 137 (5.39) 50.5 (1.99)
G1¼ 61 (2.40) 44.5 (1.75)
重量 材料
1.5...1.8 kg (3.3...4.0 lbs)，取决于具体型号 EPDM ( 密封圈 ) PVC PP ( 法兰 )
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过程连接
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过程连接示意图 A B C D E F G1¼ 外螺纹， PVC ANSI 1" 法兰， PVC DN 25 胶合端口， PVC D20 软管， PVC Rp ¾" 内螺纹， PVC NPT ¾" 内螺纹， PVC
配套传感器
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Endress+Hauser中国销售中心
深圳市百合顺电子科技有限公司 地址：深圳市龙岗区丹竹头仙桃源工业区3栋601 电话：0755-29309336 传真：0755-29309339 邮箱：955855@ 网站：
TI00495C/28/zh/02.12 FM 9.0
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CYA251，带清洗喷头 1 清洗喷头
传感器适配接头
CYA251 的传感器适配接头 • 适配接头，用于安装下列传感器 • 订货号：71144333 ( 适用于 COS61D、 COS61、 COS31 传感器 ) • 订货号：71144334 ( 适用于 COS41、 COS51D 传感器 ) • 订货号：71144335 ( 适用于 CUS51D 传感器 ) • 订货号：71144336 ( 适用于 CUS31、 CUS41 传感器 ) • 订货号：71144337 ( 适用于 CAS51D 传感器 (2 mm 光程 )) • 订货号：71144338 ( 适用于 CAS51D 传感器 (8 mm 光程 )) • 订货号：71144339 ( 适用于 CAS51D 传感器 (40 mm 光程 ))
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Flowfit CYA251
机械结构
外形尺寸
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外形尺寸示意图
传感器
CAS51D 2 mm 468 (18.43)
CAS51D 8 mm 471 (18.54)
CAS51D 40 mm 477 (18.78)
COS51D COS41 284 (11.18)
压力 - 温度曲线
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压力 - 温度曲线
流量
取决于所使用的传感器和传感器属性。 典型值： 最小值： 最大值： 溶解氧传感器：约 200 l/h (53 gal/hr) 浊度传感器和紫外法 (UV) 传感器：约 100 l/h (26.5 gal/hr) 取决于测量参数，但是必须选择有代表性的参数。 流量大于 300 l/h (80 gal/hr) 时，不建议使用。
p1
p2
p1 > p2
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连接示意图：在包含旁通管和孔板的主管道中安装 ( 安装支架 90 ° 旋转，进水口在底部 )
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Flowfit CYA251
无需采取其他措施增大主管道的旁通管管道压力。
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连接示意图：带开放式出水口
流通式安装支架的进水口和吹水口连接始终相同。 流通式安装支架未指定进水口侧和出水口侧。因此，可以针对特定应用进行选择。 壁式安装支架