mkt复习资料 (2)

mkt复习资料 (2)
mkt复习资料 (2)

第一章

1. Marketing: A social and managerial process where by individuals and groups obtains what they need and want through creating and exchanging products and value with others.

Needs需要:A state of felt deprivation. (Include physical need, social needs and individual need) Wan t欲望: The form taken by a human need as shaped by culture and individual personality Just like Chinese wants rice, the Japanese sushi, and the Korean kimchi.

Demands需求:Human wants that are backed by buying power (at Disney theme parks, spends a day touring the park in a Mickey, Minnie, Winnie the Pooh, or other character costume.)

2. Marketing management orientations营销管理导向

●Production concept生产观念:The idea that consumers will favor products that are

available and highly affordable.

●Product concept产品观念:

The idea that consumers will favor products that offer the most quality, performance, and features and that the organization should therefore devote its energy to making continuous product improvement.

●Selling concept销售观念:the idea that consumers will not buy enough of the organization’s

products unless the organization undertakes a large-scale selling and promotion effort.

●Marketing concept营销观念:the marketing management philosophy that holds that

achieving organization goals depends on determining the needs and wants of target markets and delivering the desired satisfactions more effectively and efficiently than competitors do.

●Societal marketing concept社会营销观念: the idea that the organization should determine

the needs, wants, and interests of target markets and deliver the desired satisfactions more effectively and efficiently than do competitors in a way that maintains or improves the consumer’s and society’s well-being.

第二章

1. Mission statement使命陈述: a statement of the organization’s purpose-what it wants to accomplish in the larger environment.

It’s time to ask: What is our business? Who is the customer? What do consumers value? What should our business be?

Mission statements should be market oriented

Mission should be realistic

Missions should also be specific

Mission should fit the market environment

The organization should base its distinctive competencies

Finally, mission statements should be motivating

2. The BCG growth-share matrix波士顿成长-份额矩阵

Stars明星类are high-growth, high-share businesses or products.

Cash cows金牛类are low-growth, high –share business or products.

Question marks问题类are low-share business units in high-growth markets.

Dogs瘦狗类are low-growth, low share business and products.

3.价值链和价值传递网络区别

Value chain价值链: The series of departments that carry out value-creating activities to design, product, market, deliver, and support a firm’s products.

Value-delivery network价值传递网络:The network made up of the company, suppliers, distributors, and ultimately customers who ―partner‖ with each other to improve the performance of the entire system.

比较To create customer value, the firm must look beyond its own value chain and into the value chains of its supplies, distributions, and, ultimately, customers. More companies today are partnering with the other members of the supply chain to improve the performance of the customer value-delivery network.

Increasingly in today’s marketplace, competition no longer takes place between individual competitors. Rather, it takes place between the entire value=delivery networks created by these competitors.

5. Market segmentation市场细分: Dividing a market into distinct groups of buyers who have distinct needs, characteristics, or behavior and who might require separate products or marketing mixes.

Market segment细分市场:A group of consumers who respond in a similar way to a given set of marketing efforts.

Target marketing目标市场:the process of evaluating each market segment’s att ractiveness and selecting one or more segments to enter.

Market positioning市场定位: Arranging for a product to occupy a clear, distinctive, and desirable place relative to competing products in the minds of target consumers.

第四章

Economic environment经济环境:Factors that affect consumer buying power and spending patterns.

There are four types of industrial structures产业结构类型: subsistence economies like Laos (few opportunities for marketers); raw –material-exporting economics like Brunei (oil), with good markets for equipment tools, supplies, and luxury goods for the rich; industrializing economies, like India and china, where a new rich class and a growing middle class demand new types of goods; and industrial economics like Japan , which are rich markets for all sorts of goods.

第六章

2.Model of buyer behavior消费者行为模型

Marketing and other stimuli Buy’s black box buyer responses Marketing other Buyer buyer product choice Product economic characteristics decision process brand choice

Price technological dealer choice Place political purchase timing Promotion cultural purchase amount 3.Social classes社会阶层:Relatively permanent and ordered divisions in a society whose members share similar values, interests, and behaviors. Social classes show distinct product and

brand preferences in areas such as clothing, home furnishings, leisure activity, and automobiles.

为什么这点加以关注Marketers are interested in social class because people within a given social class tend to exhibit similar buying behavior.

4. Opinion leaders意见领袖: Person within a reference group who, because of special skills, knowledge, personality, or other characteristics, exerts influence on others.

7章

Systems selling系统销售: Buying a packaged solution to a problem from a single seller ,thus avoiding all the separate decisions involved in a complex buying situation.

(步骤) First ,the supplier sells a group of interlocking products .For example ,the supplier sells not only glue, but also applicators and dryers.

Second, the suppliers sell a system of production, inventory control, distribution, and other services to meet the buyer's need for a smooth-running operation.

Evaluating market segments 细分市场评估:p201

1.Evaluating因素Three factors: segment size and growth, segment structural attractiveness ,and company objectives and resources.

●First collect and analyze date on current segment sales, growth rates, and expect profitability

for various segments. It will be interested in segments that have the right size and growth characteristics.

● A segment is less attractive if it already contains many strong and aggressive competitors.

The relative power of buyers also affects segment attractiveness. A segment may be less attractive if it contains powerful suppliers who can control prices or reduce the quality or quantity of ordered goods or servicers.

●If the company lacks the strengths needed to compete successfully in a segment and cannot

readily obtain them, it should not enter the segment. Even if the company possesses the required strengths, it needs to employ skills and resources superior to those of the competition to really win I a market segment.

第九章

1. Levels of product and services: 产品和服务的层次

Three levels: the most basic level is the core benefit, which addresses the question. What is the buyer really buying? At the second level, product planners must turn the core benefit into an actual product. They need to develop product and services features, design, a quality level, a brand name, and packaging. Finally, product planners must build an augmented product around the core benefit and actual produce by offering additional consumer services and benefits.

2. Branding 品牌

A name, term, sign, symbol or design, or a combination of these, Intended to identify the goods or services of one seller or group of sellers and to differentiate them from those of competitors. For example, most consumers would perceive a bottle of White Linen perfume as a high-quality, expensive product. But the same perfume in an unmarked bottle would likely be viewed as lower in quality, even if the fragrance were identical.

3. Product line decisions产品线决策

A product line is a group of products that are closely related because they function in a similar manner, are sold to the same customer groups, are marked through the some types of outlets, or fall within given price ranges. Sony Ericsson produces several lines of telecommunications products

A company can lengthen its product line in two ways 公司可以通过两种方法延长其产品线

(1) Product line stretching occurs when a company lengthens its product line beyond its current range, the company can stretch its line down ward, upward, or both ways.

(2) An alternative to product line stretching is product line filling-addend more items within the present range of the line.

There are several reasons for product line filling: reaching for extra profits satisfying dealer, using excess capacity, being the leading full-line company, and plugging holes to keep out competitors. For example: Sony filled its Walkman line by addend solar-powered and waste proof Walkman, an ultralight model that attachés

4. Product mix decisions 产品组合决策

定义:The set of all product lines and items that a particular seller offers for sale.

A company’s product mix has four important dimensions: width, length, depth ,and consistency. Product mix width refers to the number of different product lines the company carries. For example, Procter& gamble markets a fairly wide product mix consisting of 250 brands organized into many product lines.

●Product mix length refers to the total number of items the company carries within its product

lines. P&G typically carries many brands within each line, it sells seven laundry detergents, six hand soap, five shampoos and four dishwashing detergents

●Product line depth refers to the number of versions offered of each, product in the line.

Crest toothpaste comes in 13 variet8es.

●Finally, the consistency of the product mix refers to how closely related the various product

lines are in end use, production requirements, distribution channels, or some other way.

5. Brand equity品牌资产

The positive differential effect that knowing the brand name has on customer response to the product or service.

重要性A brand with strong brand equity is a very valuable asset. High brand equity provides a company with many competitive advantages. A powerful brand enjoys a high level of customer brand awareness and loyalty. Because consumers expect stores to carry the brand , the company has more leverage in bargaining with resellers. Because the brand name carries high credibility, the company can more easily launch line and brand extensions, as when Coca-Cola leveraged its well-known brand to introduce Diet Coke. Above all, a powerful brand offers the company some defense against fierce price competition.

6. Brand positioning品牌定位

Major brand strategy decisions主要的品牌战略决策

Brand positioning Brand name selection Brand sponsorship Brand development Attributes selection manufacturer’s brand Line extensions

Benefits → protection → private brand → Brand extensions Beliefs and values Licensing Multibrands

Co-branding New brands

7. Brand development品牌发展

Line extension产品线延伸Using a successful brand name to introduce additional items in a given product category under the same brand name, such as new flavors, forms, colors, added ingredients, or package sizes.

Brand extension品牌延伸Using a successful brand name to launch a new or modified product

in a new category.

Multibands多品牌: companies often introduce additional brands in the same category.

New brands新品牌: A company may create a new brand name when it center a new product category for which none of the company’s current brand names is appropriate.

Brand development strategies 品牌发展战略

Product Category

Existing New

Brand Existing line extension brand extension

Name New Multibrands New brands.

8. Services marketing and characteristic s

Service intangibility服务无形化

A major characteristic of services -----they cannot be seen, tasted, felt, heard, or smelled before they are bought. Services cannot be seen, tasted, felt, heard, or smelled before purchase. Service inseparability 服务不可分性

they are produced and consumed at the same time and cannot be separated from their providers, whether the providers are people or machines. Services cannot be separated from their providers. Service variability服务易变性

Their quality may vary greatly, depending on who provides them and when, where, and how. Quality of services depends on who provides them and when, where, and how.

Service perishability服务易逝性

They cannot be stored for later sale or use. Services cannot be stored for later sale or use.

10章

Product life cycles产品周期的四个阶段

Introduction stage:The product life-cycle stage in which the new product is the first distribution and made available for purchase.

Growth stage:The product life- cycle stage in which a product’s sales start climbing quickly. Maturity stage:The stage in the product life cycle in which sales growth slows or levels off. Decline stage:The product life-cycle stage in which a product’s sales decline.

12章

New-product Pricing Strategies新品定价策略p302

1.Market-skimming Price市场撇制定讲价:Setting a high price for a new product to skim maximum revenues layer by layer from the segments willing to pay the high price; the company makes fewer but more profitable sales.

2.Market-penetration Pricing市场渗透定价: Setting a low price for a new product in order to attract a large number of buyers and a large market share.

Product mix pricing strategies产品组合定价战略p303

1.Product line pricing产品线定价: setting the price steps between various products in a product line based on cost differences between the products, customer evaluations of different features, and competitors' prices.

例子In many industries, sellers use well-established price points for the products in their line. Thus, men's clothing stores might carry men's suits at 3 price levels: $185, $325, and $495. The customer will probably associate low-, average-,and high-quality suits with the three price points. Even if the three prices are raised a little, men normally will buy suits at their own preferred price

points. The seller's task is to establish perceived qualities that support the price differences.

2.Optional-product pricing可选择产品定价: the pricing of optional or accessory products along with a main product.

例子A car buyer may choose to order power windows, cruise control, and an extended warranty.

3.Captive-product pricing捆绑定价:setting a price for products that must be used along with a main product, such as blades for a razor and film for a camera.

例子Nintendo sells its game consoles at low price and makes money on video game titles. While Nintendo's margins on its consoles run a mere one percent to five percent, margins on its game cartridges run close to 45 percent. Video game sales contribute more than half the company's profits.

4.By-product pricing副产品定价: setting a price for by-products in order to make the main product's price more competitive.

例子Malaysia's Golden Hope Plantations used to burn old rubber trees after their productive life.Now, it cuts them down and use the wood to supply the fast-growing markets for rubber wood furniture, parquet flooring, and particle and medium-density fiber boards.

5.Product bundle pricing产品捆绑定价: combining several products and offering the bundle at

a reduced price.

例子Theaters and sports teams sell season tickets at less than the cost of single tickets. Segmented pricing差别定价:selling a product or service at two or more prices, where the difference in prices is not based on differences in costs.(4个)

1.customer-segment pricing

2.product-form pricing

3.location pricing 4 .time pricing

Price Changes: p311

降价initiating price cuts:one such circumstance is excess capacity. Another situation leading to price changes is falling market share in the face of string price competition .A company may also cut prices in a drive to dominate the market through lower costs .

涨价initiating price increases:A major factor in price increases is cost inflation .Rising costs squeeze profit margins and lead companies to pass cost increases along to customers .Another factor leading to price increases is over demand :When a company cannot supply all its customers' needs ,it can raise its price ,ration products to customers ,or both .

价格反映战略(应对对手涨价4个方法):p314

1. It could reduce its price to match the competitor's price.

2. The company might maintain its price but raise the perceived quality of its offer.

3. The company might improve quality and increase price, moving its brand into a higher-price position.

4. The company might launch a low-price "fighting brand"---adding a lower-price item to the line or creating a separate lower-price brand.

第十三章

Number of Channel level:

Direct marketing channel:A marketing channel that has no intermediary

Indirect marketing channel:Channel containing one or more intermediary levels.

Channel conflict:Disagreement among marketing channel members on goals and roles-who should do what and for what rewards.

Vertical marketing system(VMS):A distribution channel structure in which

producers,wholesalers,and retailers act as a unified system.One channel member owns the others, has contracts with them,or has so much power that they all cooperate.

Corporate VMS公司式垂直营销系统:A vertical marketing system that combines successive stages of production and distribution under single ownership--channel leadership is established through common ownership.

Contractual VMS合同式: A vertical marketing system in which independent firms at different levels of production and distribution join together through contracts to obtain more economies or sales impact than they could achieve alone.

Administered VMS管理式:A vertical marketing system that coordinates successive stages of production and distribution, not through common ownership or contractual ties, but through the size and power of one of the parties.

Horizontal marketing system水平营销系统: A channel arrangement in which two or more companies at line level join together to follow a new marketing opportunity.

Number of Marketing Intermediaries中间商数量:

Intensive distribution密集型分销: Stocking the product in as many outlets as possible. For example, toothpaste, candy, and other similar items are sold in millions of outlets to provide maximum brand exposure and consumer convenience. Coca-cola and other consumer goods companies distribute their products in this way.

Exclusive distribution专营性分销: Giving a limited number of dealers the exclusive right to distribute the company’s products in their territories. For example, Benz dealers are few and far between—even large cities may have only one dealer. By granting exclusive distribution, Benz gains stronger distributor selling support and more control over dealer prices, promotion, credit, and services.

Selective distribution选择性分销:the use of more than one, but fewer than all, of the intermediaries who are willing to carry the company’s products. For example, national, Mitsubishi, and Samsung sell their major appliances through dealer networks and selected large retailers. By using selective distribution, they do not have to spread their efforts over many outlets, including many marginal ones. They can develop good working relationships with selected channel members and expect a better- than-average selling effort.

第十六章

Advertising objective广告目标: A specific communication task to be accomplished with a specific target audience during a specific period of time.

3种advertising objective: informative, persuasive reminder adverting.

Consumer Promotion Tools销售促进9种方法:

Samples样本: A small amount of a product offered to consumers for trial,

Coupons优惠券: Certificate that gives buyers a saving when they purchase a specified product, Cash refunds offers现金折扣:Offer to refund part of the purchase price of a product to consumers who send a "proof of purchase" to the manufacturer,

Price packs特价包:Reduced price that is marked by the producer directly on the label or package,

Premiums赠品: Good offered either free or at low cost as an incentive to buy a product, Advertising specialties广告礼品: Useful article imprinted with an advertiser's name, given as a gift to consumers,

Patronage rewards光顾奖励: Cash or other award for the regular use of a certain company's products or services,

Point-of-purchase promotions售点陈列: Display and demonstration that takes place at the point of purchase or sale,

Contests, sweepstakes, games竞赛抽奖和游戏: Promotional events that give consumers the chance to win something--such as cash, trips, or goods--by luck or through extra effort.

荧光标记二抗的选择

荧光标记二抗的选择 荧光标记二抗的选择-FITC/Rhodamine/Texas Red/Cy/PE/AMCA 一般来讲,耦联到二抗上的探针主要有酶(辣根过氧化酶HRP和碱性磷酸酶AP 或其衍生物APAAP,PAP),荧光基团(FITC, RRX, TR, PE, Rhodamine)和生物素。选用哪种探针的二抗主要取决于具体的实验。对于Western Blot和ELISA,最常用的二抗是酶标二抗;而细胞或组织标记实验(细胞免疫化学,组织免疫化学,流式细胞术)中通常使用荧光标记的二抗。如果想要更大程度的放大检测信号,可以使用Biotin/Avidin检测系统。其中荧光素是具有光致荧光特性的染料,荧光染料种类很多,目前常用于荧光标记二抗有以下几种: 【异硫氰酸荧光素-Fluorescein Isothiocyanate (FITC)荧光标记二抗】 FITC纯品为黄色或橙黄色结晶粉末,易溶于水和酒精溶剂。FITC分子量为389.4,最大吸收光波长为490~495nm,最大发射光波长为520~530nm,呈现明亮的黄绿色荧光。FITC在冷暗干燥处可保存多年,是目前应用最广泛的荧光素。由于FITC是小分子化合物,每一个抗体可标记几个FITC分子,IgM通常用小分子的荧光素标记,如FITC、Cy3/5、Texas Red等。FITC荧光二抗主要优点是人眼对黄绿色较为敏感,通常切片标本中的绿色荧光少于红色。然而FITC的最大缺点是淬灭快,因此要和抗淬灭剂一起使用。 【四甲基异硫氰酸罗丹明-Tetramethyl Rhodamin Isothiocyanate(TRITC),Rhodamine Red-X(RRX), Texas Red(TR)荧光标记二抗】 这些罗丹明的衍生物耦联基团具有不同的吸收波长(550, 570, 596nm)和最大发 射波长(570, 590, 620nm)。尽管TRITC经常和FITC一起在双标记实验中使用,使用RRX和TR可以得到更好的颜色区分。在使用装有氪氩灯的激光共聚焦扫描显微镜作三标记的实验时,RRX尤其有用,可以和Cy2(或者FITC)和Cy5一起使用,因为RRX的发射波长在Cy2和Cy5中间,而且和这两者覆盖都很少。氪氩灯激发光为488nm,598nm和647nm,分别是Cy2(FITC), RRX和Cy5的理想激发波长。因为FITC和PE可以被氩灯的488nm波长激发, 在流式细胞仪中用FITC作双标,另一种用藻红蛋白(PE)耦联基团要比罗丹明好。TRITC为罗丹明的衍生物,呈紫红色粉末,较稳定。最大吸收光波长为550nm,最大发射光波长为620nm,呈现橙红色荧光,与FITC的绿色荧光对比鲜明,可配合用于双重标记或对比染色。因其荧光淬灭慢,也可用于单独标记染色。 【菁类染料-Cyanine dyes(Cy2, Cy3, Cy5)】 Cy2耦联基团激发波长为492nm,发光为波长510nm的绿色可见光。Cy2和FITC 使用相同的滤波片。由于Cy2比FITC在光下更稳定。要避免使用含有磷酸化的苯二胺的封片剂,因为这种抗淬灭剂和Cy2反应,在染色片储存后会导致荧光微弱和扩散。Cy3和Cy5比其他的荧光团探针要更亮,更稳定,背景更弱。Cy3耦联基团激发光的最大波长为550nm,最强发射光为570nm。因为激发光和发射光波长很接近TRITC, 在荧光显微镜中,可使用和TRITC一样的滤波片。 Cy3在氩光灯(514nm或528nm)下可以被激发出50%的光强,在氦氖灯(543nm)或者汞灯(546nm)下则约75%。Cy3可以和荧光素一起作双标。Cy3还可以和Cy5一起在共聚焦显微镜实验中作多标记。Cy5耦联基团的激发波长最大650nm,发光波长最大670nm。在氪氩灯(647nm)下它们可被激发出98%的荧光,在氦氖灯下(633nm)为63%。Cy5可以和很多其他的荧光基团一起用在多标记的实验中。由于它的最大发射波长在670nm,Cy5很难用裸眼观察,而且不能用汞灯作理想

教你如何选择二抗

*教你如何正确选择二抗 通常情况下,某一特定的实验中可能同时有几种二抗可供选择,如何能选择到最适合该实验的二抗,需要综合以下几个方面进行考虑: ●一抗的物种来源 一抗是在什么物种来源的,相应的二抗也要是抗该物种的抗体。例如,如果一抗是在小鼠体内制备的,那么你就应当选择抗小鼠的二抗,如果一抗是在兔体内制备的,那么二抗就应当是抗兔的抗体。至于二抗本身是在什么动物中制备的对二抗的质量并无明显的影响。 ●一抗是属于哪个类或亚类 二抗需与一抗的类别或亚类相匹配。这通常是针对单克隆抗体而言。多克隆抗体主要是IgG类免疫球蛋白,因此相应的二抗就是抗IgG抗体. 单克隆抗体的类别及亚类通常会在产品列表中列出,如果你的一抗是小鼠IgM,那么相应的二抗就应当是抗小鼠IgM,或是抗小鼠IgG抗体。 如果单克隆一抗是小鼠IgG的某一亚类(IgG1,IgG2a,IgG2b,IgG3),那么几乎所有的抗小鼠IgG都可以与之结合,或者你也可以选择专门针对这一亚类的二抗,例如,如果你的一抗是小鼠IgG1,那么你可以选择抗IgG1的二抗,此种抗体在双标记实验中尤其适合。 如果你不知道一抗是哪一类别或亚类,那么抗小鼠IgG是一个不错的选择,因为此种抗体可以识别大多数类型的IgG免疫球蛋白。

二抗的特异性 下面总结了几种具有不同特异性的二抗:针对整个抗体分子(H+L)具有特异性:如抗IgG(H+L),此类抗体既可以与抗体的重链结合也可以与轻链结合,即与抗体分子的Fc,F(ab’)2/Fab部分(见图5)均可反应,抗IgG(H+L)也可以与其他免疫球蛋白家族反应(如IgM和IgA),因为所有的免疫球蛋白都具有相同的轻链(kappa链或lambda 链)。 针对Fab片段具有特异性:这类抗体与重链轻链均可以结合,由于它们可以与轻链反应,它们同时也已和具有相同轻链的其他种类的免疫球蛋白反应。 这对Fc片段或重链具有特异性:这类抗体和重链的Fc部分反应,一次它们是类别特异性的(即gamma链特异性抗体只与IgG反应,mu链特异性抗体只识别IgM,依此类推)。 轻链(kappa,labmda)特异性:与所有类别的抗体反应,因为所有类别的抗体均具有相同的lambda链或kappa链。 经过吸附处理的二抗(cross-absorbed):不同来源的免疫球蛋白含有相似的结构,抗一个物种的抗体可能与其他物种发生交叉反应,一次有些二抗经过了动物或人类IgG吸附处理,以减少非特异性背景。例如,如果是小

Westernblot一抗和二抗的选择讲解学习

W e s t e r n b l o t一抗和二抗的选择

精品文档 Westernblot一抗和二抗的选择 抗体分类:根据重链恒定区的血清学类型,可将抗体分为IgM,IgG,IgA,IgD,IgE 五类,它们的重链分别为mu, gamma, alpha, delta, epsilon链。在上述每一类别中,按重链构造上的变异又可分为几个亚类,例如人的IgG可分为IgG1,IgG2,IgG3,IgG4四个亚类。轻链分为两种类型,kappa链和lambda链,但每种抗体中只存在一种类型的轻 链。二抗:二抗是在其它宿主体内制备的能与一抗或一抗片段结合的抗体,上面通常连有酶或荧光素等标签。由于二抗所具备的优点使得其在免疫学实验中得以应用广泛,如western blot(通过与特异性抗体结合来鉴定蛋白质),ELISA(以耦联有酶的抗体或抗原为标记来检测特异性的蛋白质,尤其是相应的抗原或抗体),免疫组织化学(检测组织中的特异性抗原),免疫细胞化学(通过免疫学方法检测细胞的抗原组成),流式细胞术(通过检测激光所激发荧光来鉴定分离不同类型的细胞)及免疫沉淀(通过抗原与抗体的特异性结合作用来分离相应抗原)。二抗针对某一特定物种(如小鼠)的所有抗体均具有特异性,因而使用标记的二抗可以免去对每一个一抗进行标记,大大节省了时间和费用;此外,一个一抗分子可以同时结合几个二抗分子,从而使信号大大增强,提高了实验灵敏度。 二,如何选择二抗——根据一抗种属及类型选择合适的二抗 广义上是指专门和进行特异性反应和结合的抗体,在免疫学反应中,经常需要针对试验选择不同的二抗,**捷能为您的科研工作提供最适合和最全面的二抗产品。检测任何目的靶蛋白都有不止一种抗体可供选择,同时在后继试验中也会有不同的检测方案,因此在选择二抗的时候要综合考虑一抗的类型及后继检测方案的要求,一般来说,选择合适的二抗需要从下面几个方面考虑:【一抗的种属来源】 二抗应选用与使用的一抗相同的物种来源,例如:如果你的一抗是小鼠源的单克隆抗体,二抗则选抗小鼠的二抗(山羊抗小鼠或者兔抗小鼠等均可);如果一抗是从兔血清里制备的兔源多克隆抗体,则相应的二抗需要选择抗兔的二抗。即根据一抗的物种来源选择相应的抗该物种的二抗。**捷能为您提供最全的抗不同种属的原装进口二抗,包括抗小鼠、大鼠、兔、山羊、绵羊、人、豚鼠、猪、马、牛、鸡、鸭等二抗。【一抗的类别亚型】 二抗需与一抗的类别或亚类相匹配。这通常是针对单克隆抗体而言。多克隆抗体主要是IgG类免疫球蛋白,因此相应的二抗就是抗IgG抗体。其中单克隆抗体的类别及亚类通常会在产品说明书中都会有描述,如果你的一抗是小鼠IgM,那么相应的二抗就应当是抗小鼠IgM。如果单克隆一抗是小鼠IgG的某一亚类(IgG1,IgG2a,IgG2b,IgG3),那么几乎所有的抗小鼠IgG都可以与之结合,或者你也可以选择专门针对这一亚类的二抗,例如,如果你的一抗是小鼠IgG1,那么你可以选择抗IgG1的二抗,此种抗体在双标记实验中尤其适合。在不清楚一抗为何种类/亚类的情况下,可以选用抗相应物种IgG。**捷能为您提供通用的IgG(H+L)二抗,或者特异性只结合IgM的Anti IgM (mu) Antibody、IgA的Anti IgA (alpha-Antibody、IgE的Anti IgE (epsilon) Antibody等。【二抗的种属来源】 一般来说,不同的种属来源与二抗的质量没有必然的联系,来源于山羊的二抗与来源于驴的二抗在一般的实验里没有太多的差别。然而在一些特殊的实验里,如双标实验里,如果其中一个一抗是山羊来源的,一个是小鼠来源的,则相应的二抗分别要抗山羊和抗小鼠的 收集于网络,如有侵权请联系管理员删除

如何选择二抗

如何选择二抗 抗体简介 抗体:是高等动物在抗原物质的刺激下由浆细胞产生的一类能与相应抗原发生特异性结合的免疫球蛋白。抗体通常存在于血清合淋巴(组织液)中。 单克隆抗体及多克隆抗体:由单一克隆的细胞(通常是单一克隆的杂交瘤细胞)所产生的识别某一抗原表位的单一抗体;多克隆抗体是由多种细胞分别接触相应抗原而产生的多种抗体的总和,多克隆抗体抗原识别抗原的多个表位。 抗体结构:抗体是由四条肽链构成的“Y”形对称的分子,包括两条重链和两条轻链。在“Y”的尖端部位的氨基酸顺序在不同的抗体分子中差别很大,是负责与抗原进行特异性结合的区域,称为可变区,余下的区域则称为恒定区。 抗体分类 抗体分类:根据重链恒定区的血清学类型,可将抗体分为IgM,IgG,IgA,IgD,IgE五类,它们的重链分别为mu, gamma, alpha, delta, epsilon链。在上述每一类别中,按重链构造上的变异又可分为几个亚类,例如人的IgG可分为IgG1,IgG2,IgG3,IgG4四个亚类。 轻链分为两种类型,kappa链和lambda链,但每种抗体中只存在一种类型的轻链。 二抗:二抗是在其它宿主体内制备的能与一抗或一抗片段结合的抗体,上面通常连有酶或荧光素等标签。由于二抗所具备的优点使得其在免疫学实验中得以应用广泛,如western blot(通过与特异性抗体结合来鉴定蛋白质),ELISA(以耦联有酶的抗体或抗原为标记来检测特异性的蛋白质,尤其是相应的抗原或抗体),免疫组织化学(检测组织中的特异性抗原),免疫细胞化学(通过免疫学方法检测细胞的抗原组成),流式细胞术(通过检测激光所激发荧光来鉴定分离不同类型的细胞)及免疫沉淀(通过抗原与抗体的特异性结合作用来分离相应抗原)。二抗针对某一特定物种(如小鼠)的所有抗体均具有特异性,因而使用标记的二抗可以免去对每一个一抗进行标记,大大节省了时间和费用;此外,一个一抗分子可以同时结合几个二抗分子,从而使信号大大增强,提高了实验灵敏度。 二抗可以耦联有几种不同的标记,可以是酶,荧光素,或生物素。 抗体特异性产品 整个分子(H+L) 特 Affinity Purified Anti-BOVINE IgG (H&L) (GOAT) 异性抗体 Peroxidase Conjugated Anti-BOVINE IgG (H&L) (GOAT) Fluorescein Conjugated Affinity Purified Anti-BOVINE IgG (H&L) (GOAT) Fab 特异性抗体Peroxidase Conjugated Anti--HORSE IgG F(ab)2 (GOAT)

Jackson ImmunoResearch 公司二抗选用指南

Jackson ImmunoResearch 公司二抗选用指南 (此部分内容来源于Jackson ImmunoResearch Laboratories Inc网站,如有疑问,请参考原网站) 亲和层析法纯化的二抗 用亲和层析法纯化的抗体是指,利用耦联到琼脂糖凝胶上的 抗原将抗血清中的抗体亲和层析下来。我们采用一种特有的 连续的洗脱过程将抗体从固态抗原上分离出来。我们提供的 未标记的亲和层析法纯化的抗体是不含有稳定剂和防腐剂的 无菌液体。耦联标记物的抗体是冻干粉,里面含有稳定剂和 防腐剂,但是过氧化物酶标记的抗体里面不含防腐剂。 (下面内容中的页码指的是Jackson 2006年原版目录中的页 码) 亲和层析纯化抗体的选择和定位 第一步:选择Whole IgG(8-17页)、F(ab’)2片段(18-22页),或者Fab片段(23-25)抗体。 我们提供三种亲和纯化的抗体:Whole IgG,F(ab’)2片段,Fab片段。 Whole IgG(8-17页)抗体是从抗血清中分离出来的完整的分子。这些抗体有一个Fc部分和两个与抗原结合的Fab部分,因此是二价的。Whole IgG抗体适用于多数情况(见F(ab’)2片段、Fab片段的特例),也是性价比最高的。 F(ab’)2片段(18-22页)抗体是用胃蛋白酶消化IgG去除Fc部分,剩下两个靠二硫键连接的结合抗原的Fab部分(依然是二价的)。这些抗体用在特定的情况中,例如避免抗体与Protain A 或G 结合,或者与有Fc受体的活细胞结合。但是,如果一抗是whole IgG,不管二抗是哪种形式都可能出现结合Fc受体。这时,为了阻止抗体结合到Fc受体上,可以在4度将细胞预培养于含有叠氮钠和二抗来源物种正常血清的缓冲液中。

WB检测抗体如何选择(下)

WB检测抗体如何选择 WB检测之二抗 二抗选择 二抗是用一抗免疫动物制备的抗一抗的抗体,选择范围较窄,基本是商品化的,在不考虑二抗修饰类型的前提下,二抗的选择需要遵循以下原则:二抗来源种属、一抗来源种属和抗原来源种属互不相同。例如如果研究对象是人的蛋白X,选择了鼠抗X一抗,则可选择兔抗鼠二抗或者羊抗鼠二抗。 二抗选择要考虑的问题有: ①对单抗类一抗,二抗需与一抗的类别或亚类相匹配(多抗类一抗主要是IgG类免疫球蛋白故相应二抗就是抗 IgGF(ab’)2二抗,避免各种Ig保守结构域导致的交叉反应。然而各种Ig仍具有保守的轻链结构域,因此还是有一定程度的交叉反应。 WB检测之内参抗体 Western Blot检测方法不仅能检测靶蛋白存在与否,还能比较不同条件下或者不同组织中靶蛋白的相对表达量,可作为蛋白表达水平最直接的证据。在WB实验中有众多不确定因素影响靶蛋白表达水平的测定,如方法学本身性质、操作误差等。为准确衡量靶蛋白表达水平,需要精确一致的上样量,使用内参的意义即是保证上样量的一致。内参即内部参照(Internal Control),对哺乳动物细胞一般指管家基因的表达蛋白(Housekeeping Proteins)。此类蛋白在各组织和细胞中的表达相对恒定,可作为检测蛋白表达水平变化的参照物。当内参条带亮度基本一致时,基本可以确定上样量是一致的,通常采用比较靶蛋白和内参蛋白条带亮度比值来进行相对定量。 WB实验中涉及到的蛋白-抗体结合是一个复杂的活性生物大分子间的相互作用,众多因素都会影响实验的结果。例如时间、温度、浓度、离子强度等,在实验设计时就需要有严谨的对照方案,通过对照就容易判断问题所在,严谨的WB实验中需要设立蛋白分子量标准、空白载体对照、未诱导对照、已知量标准物正对照和内参。 内参使用方法 1.标记内参:酶标的内参抗体可避免二抗结合总蛋白中某些免疫球蛋白产生的杂带,使用时只需在二抗孵育时加入酶标内参抗体,按照正常操作即可。 2.普通内参:当靶蛋白分子量与所选内参分子量相差不大时,可先对靶蛋白进行显色检测,然后用Strip缓冲液洗掉膜上抗体,重新对内参蛋白进行显色检测。当靶蛋白分子量与所选内参分子量相差较明显时,可对转膜预染,根据蛋白质分子量将膜剪为两部分并分开进行靶蛋白和内参蛋白的显色检测。 内参抗体选择 常用的蛋白内参有GAPDH和β-actin或β-tubulin,内参的选择原则是选择与靶蛋白分子量相差5KD以上的内参,对不同的样本则需要根据实际情况进行选择。 1.根据物种选择 哺乳动物来源的组织或细胞样本通常选择β-actin、β-tubulin、GAPDH、Lamin B、Histone H3、Na+/K+-ATPase等。植物来源的样本常选择plant actin、Rubisco等。对于其他研究稀少的物种可参考报导的文献进行选择。以GAPDH为例,抗GAPDH单抗(ABGENT,clone 6C5)能够与非洲蟾蜍、猴、猪、羊、狗、猫、鱼、蛙、鸡、兔、小鼠、大鼠及人组织来源的GAPDH反应,但不能与酵母GAPDH反应。 2.根据组织类型选择

一抗的选择

一抗得选择 检测任何目得靶蛋白都有不止一种抗体可供选择,为缩小抗体得选择范围选中合适得抗体,需要考虑如下几种因素: 分析试验应用得类型l 样本蛋白得结构性质l 样本得种属l 抗体宿主得种类l 抗体得标记与检测l 1、分析试验应用得类型 一般抗体说明书都列出该抗体经试验验证过适用于何种分析类型,如:可以应用于WBIHCICC ELASA分析等,如果抗体说明书没有提及得应用类型,并不意味着该抗体不适用于此种分析应用类型,而仅就是说明尚未经过此种分析试验验证,如果抗体不适用某些分析试验,则会在抗体说明书上标注出来不适于某分析试验。 2、样本蛋白得结构性质 了解样本蛋白得结构性质有助于选择最合适得抗体,至少两方面因素需要考虑 待测样本蛋白得结构域:抗体就是由各种不同免疫原免疫宿主而制备得来,其中得免疫原包括:全长蛋白、蛋白片断、多肽、全有机体(如:细菌)或细胞,抗体说明书一般都有免疫原得描述,如果打算检测得就是蛋白片断或一种特殊得同型物或蛋白全长得某一区域,则必须选择用含此片段域得免疫原制备出得抗体.如果打算用FACS流式检测活细胞得表面蛋白,则需要选择含该表面蛋白得胞外域来免疫制备得抗体。l 样本得提取或处理过程:某些抗体要求样本经过某些特殊处理,例如:许多抗体只识别还原与变性得、表位已暴露不受二级四级结构阻碍得蛋白样本,另一方面,某些抗体仅识别天然折叠状态得蛋白。当选择免疫组化得抗体时,应注意某些抗体只识别未固定得冷冻得组织,而另一些抗体则适用于无需抗原修复解交联步聚得甲醛固定石蜡包埋得组织,这些都会在抗体说明书上应用部分标示出来l 3、样本得物种 应选择物种相同或有交叉反应得抗体,抗体可能与不同物种得同种靶蛋白有交叉反应,因其氨基酸序列同源性较高,如果样本得种类未列入抗体说明书上得交叉反应种属表中,并不意味着该抗体不适用于检测该物种得蛋白,而只就是表示该物种尚未用此抗体检测验证过,应通过序列比对得方法来预测交叉反应,可应用Expasy 与NCBI BLAST来进行不同物种蛋白同源性比对。 4、一抗宿主物种得选择 一般说来,在使用偶联二抗结合无偶联物得一抗时,一抗宿主动物得物种选择较为重要,对于免疫组化而言,尽可能选择与样本不同种系物种得一抗,从而避免二抗与样本内源性免疫球蛋白产生交叉反应,例如:检测小鼠样本蛋白,则不应选择小鼠或大鼠源得一抗,最好选兔源得一抗,则二抗则可选择偶联了检测分子(酶、荧光素、生物素等)得抗兔IgG。如果选择有偶联物得一抗则不适用上述情况,除免疫组化外得其它对不含内源性免疫球蛋白样本得检测方法,则抗体宿主物种得影响不大,如对不含IgG得细胞裂解物样本得western blott ing检测,尽管如此,含有血清得组织裂解物与组织培养上清中含有免疫球蛋白,还原变性样本中含IgG,在western blot检测中则结合出现IgG分子50 and 25kDa得重链与轻链条带. 如何选择二抗-—根据一抗种属及类型选择合适得二抗 广义上就是指专门与进行特异性反应与结合得抗体,在免疫学反应中,经常需要针对试验选择不同得二抗,艾美捷能为您得科研工作提供最适合与最全面得二抗产品。检测任何目得靶

免疫荧光一抗二抗的选择及其技巧

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如何选择二抗——根据一抗种属及类型选择合适的二抗 二抗广义上是指专门和一抗进行特异性反应和结合的抗体,在免疫学反应中,经常需要针对试验选择不同的二抗,艾美捷能为您的科研工作提供最适合和最全面的二抗产品。检测任何目的靶蛋白都有不止一种抗体可供选择,同时在后继试验中也会有不同的检测方案,因此在选择二抗的时候要综合考虑一抗的类型及后继检测方案的要求,一般来说,选择合适的二抗需要从下面几个方面考虑: 【一抗的种属来源】 二抗应选用与使用的一抗相同的物种来源,例如:如果你的一抗是小鼠源的单克隆抗体,二抗则选抗小鼠的二抗(山羊抗小鼠或者兔抗小鼠等均可);如果一抗是从兔血清里制备的兔源多克隆抗体,则相应的二抗需要选择抗兔的二抗。即根据一抗的物种来源选择相应的抗该物种的二抗。艾美捷能为您提供最全的抗不同种属的原装进口二抗,包括抗小鼠、大鼠、兔、山羊、绵羊、人、豚鼠、猪、马、牛、鸡、鸭等二抗。 【一抗的类别亚型】 二抗需与一抗的类别或亚类相匹配。这通常是针对单克隆抗体而言。多克隆抗体主要是IgG类免疫球蛋白,因此相应的二抗就是抗IgG抗体。其中单克隆抗体的类别及亚类通常会在产品说明书中都会有描述,如果你的一抗是小鼠IgM,那么相应的二抗就应当是抗小鼠IgM。如果单克隆一抗是小鼠IgG的某一亚类(IgG1,IgG2a,IgG2b,IgG3),那么几乎所有的抗小鼠IgG都可以与之结合,或者你也可以选择专门针对这一亚类的二抗,例如,如果你的一抗是小鼠IgG1,那么你可以选择抗IgG1的二抗,此种抗体在双标记实验中尤其适合。在不清楚一抗为何种类/亚类的情况下,可以选用抗相应物种IgG。艾美捷能为您提供通用的IgG(H+L)二抗,或者特异性只结合IgM的Anti IgM (mu) Antibody、IgA的Anti IgA (alpha-Antibody、IgE的Anti IgE (epsilon) Antibody等。 【二抗的种属来源】 一般来说,不同的种属来源与二抗的质量没有必然的联系,来源于山羊的二抗与来源于驴的二抗在一般的实验里没有太多的差别。然而在一些特殊的实验里,如双标实验里,如果其中一个一抗是山羊来源的,一个是小鼠来源的,则相应的二抗分别要抗山羊和抗小鼠的二抗,这时候,二抗就不能选择山羊或者小鼠来源的。艾美捷有相应的驴来源的二抗,非常适合做类似双标的免疫实验。 【二抗的耦联标记】 一般来讲,耦联到二抗上的探针主要有酶(辣根过氧化酶HRP和碱性磷酸酶AP或其衍生物APAAP,PAP),荧光基团(FITC、Rhodamine、Texas Red、PE、Rhodamine、Dylight 等)、生物素、金颗粒。选用哪种探针的二抗主要取决于具体的实验。对于Western Blot和ELISA,最常用的二抗是酶标二抗;而细胞或组织标记实验(细胞免疫化学,组织免疫化学,流式细胞术)中通常使用荧光基团标记的二抗,免疫组化中也可以使用辣根过氧化酶或碱性磷酸酶标记的二抗。如果想要更大程度的放大检测信号,可以使用Biotin/Avidin检测系统。在一些荧光检测方案中,则需要选择不同的荧光标记;而金颗粒标记的二抗则更多的应用于免疫电镜中。艾美捷能为您提供上述所有不同类型标记的二抗。

如何正确选择二抗——丁香园

如何正确选择二抗二抗选择指南 2012-03-28 20:18 来源:丁香园点击次数:691 关键词:二抗单抗一抗片段基因 如何选择正确的二抗? 以下指南将帮助您选择最适合该应用的二抗。 常见问题 1.一抗的宿主物种是什么? 2.需要了解哪些有关一抗同型的内容? 3.需要酶学或荧光检测吗? 4.是否需要预吸附的二抗? 5.是否需要亲和纯化的抗体或IgG 片段? 6.有必要使用F(ab) 或F(ab')2 片段抗体吗? 7.是否需要抗IgG 重链和轻链、抗轻链或抗F(ab')2二抗? 1. 一抗的宿主物种是什么?二抗指向一抗的物种。如果使用来源于兔的一抗,则需要来源于除兔以外的其它物种的抗兔二抗。例如山羊多克隆抗兔IgG - H&L (HRP) 二抗(ab6721) 可以检测兔多克隆抗Ki67 一抗(ab15580)。 2. 需要了解哪些有关一抗同型的内容?二抗必须指向一抗同型。 多克隆一抗通常来源于兔、山羊、绵羊或驴,并且是IgG 同型。二抗通常为抗IgG 重链和轻链的抗体。单克隆一抗通常来源于小鼠、兔和大鼠。例如,如果单克隆一抗为小鼠IgG1,则需要抗小鼠IgG 或特异性较低的F(ab) 片段抗小鼠IgG。人免疫球蛋白类、亚类、型和亚型: ?类或同型:IgG (γ heavy chains), IgM (μ), IgA (α), IgE (ε), IgD (δ) ?亚类:IgG1 (γ1 heavy chains), IgG2 (γ2), IgG3 (γ3), IgG4 (γ4); IgA1 (α1), IgA2 (α2) ?型:κ light chain, λ light chain ?亚型:λ1, λ2, λ3, λ4 其它类型的反应: ?多价抗体与所有免疫球蛋白类反应 ?抗Fc 或重链(α, δ, ε, γ, and μ) 抗体仅与重链反应 ?抗F(ab) 或完整分子抗体与重链和轻链反应,而与免疫球蛋白类无关 ?由于所有免疫球蛋白类均使用相同的κ 和λ 轻链,因此抗轻链(κ 和λ)抗体与所有 免疫球蛋白类反应

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一抗的选择 检测任何目的靶蛋白都有不止一种抗体可供选择,为缩小抗体的选择范围选中合适 的抗体,需要考虑如下几种因素: 分析试验应用的类型I 样本蛋白的结构性质I 样本的种属I 抗体宿主的种类I 抗体的标记和检测I 1?分析试验应用的类型 一般抗体说明书都列出该抗体经试验验证过适用于何种分析类型,女口:可以应用于WB IHC ICC ELASA 分析等,如果抗体说明书没有提及的应用类型,并不意味着该抗体不适用于此种分析应用类型,而仅是说明尚未经过此种分析试验验证,如果抗体不适用某些分析试验, 则会在抗体说明书上标注出来不适于某分析试验。 2?样本蛋白的结构性质 了解样本蛋白的结构性质有助于选择最合适的抗体,至少两方面因素需要考虑 待测样本蛋白的结构域:抗体是由各种不同免疫原免疫宿主而制备得来,其中的免疫原包括: 全长蛋白、蛋白片断、多肽、全有机体(如:细菌)或细胞,抗体说明书一般都有免疫原的描述,如果打算检测的是蛋白片断或一种特殊的同型物或蛋白全长的某一区域,则必须选择用含此片段域的免疫原制备出的抗体。如果打算用FACS流式检测活细胞的表面蛋白,则需 要选择含该表面蛋白的胞外域来免疫制备的抗体。I 样本的提取或处理过程:某些抗体要求样本经过某些特殊处理,例如:许多抗体只识别还原 和变性的、表位已暴露不受二级四级结构阻碍的蛋白样本,另一方面,某些抗体仅识别天然 折叠状态的蛋白。当选择免疫组化的抗体时,应注意某些抗体只识别未固定的冷冻的组织,而另一些抗体则适用于无需抗原修复解交联步聚的甲醛固定石蜡包埋的组织,这些都会在抗 体说明书上应用部分标示出来I 3?样本的物种 应选择物种相同或有交叉反应的抗体,抗体可能与不同物种的同种靶蛋白有交叉反应,因其氨基酸序列同源性较高,如果样本的种类未列入抗体说明书上的交叉反应种属表中,并不意味着该抗体不适用于检测该物种的蛋白,而只是表示该物种尚未用此抗体检测验证过,应通过序列比对的方法来预测交叉反应,可应用Expasy和NCBI BLAST来进行不同物种蛋白 同源性比对。 4.一抗宿主物种的选择 一般说来,在使用偶联二抗结合无偶联物的一抗时,一抗宿主动物的物种选择较为重要,对于免疫组化而言,尽可能选择与样本不同种系物种的一抗,从而避免二抗与样本内源性免疫 球蛋白产生交叉反应,例如:检测小鼠样本蛋白,则不应选择小鼠或大鼠源的一抗,最好选 兔源的一抗,则二抗则可选择偶联了检测分子(酶、荧光素、生物素等)的抗兔IgG。如果选择有偶联物的一抗则不适用上述情况,除免疫组化外的其它对不含内源性免疫球蛋白样本 的检测方法,则抗体宿主物种的影响不大,如对不含IgG的细胞裂解物样本的western blotting 检测,尽管如此,含有血清的组织裂解物和组织培养上清中含有免疫球蛋白,还原变性样本

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