F(ab')2-Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 647

货号：A-21246

规格：250μl

价格：2787

产品类型：荧光二抗

品牌：Thermo Fisher

物种：兔

宿主：山羊

抗体亚型：IgG

荧光染料：Alexa Fluor 647

抗体类型:	荧光二抗	同型对照：	IgG
免疫原性：	Gamma Immunoglobins Heavy and Light chains	用法：	4 μg/mL (ICC)；4 μg/mL (IF)

To minimize cross-reactivity, these goat anti-rabbit IgG (H+L) divalent F(ab')2 secondary antibodies have been affinity purified and cross-adsorbed against pooled human serum, mouse serum, mouse plasmacytoma/hybridoma proteins, and purified human paraproteins. Cross-adsorption or pre-adsorption is a purification step to increase specificity of the antibody resulting in higher sensitivity and less background staining. The secondary antibody solution is passed through a column matrix containing immobilized serum proteins from potentially cross-reactive species. Only the nonspecific-binding secondary antibodies are captured in the column, and the highly specific secondaries flow through. The benefits of this extra step are apparent in multiplexing/multicolor-staining experiments (e.g., flow cytometry) where there is potential cross-reactivity with other primary antibodies or in tissue/cell fluorescent staining experiments where there are may be the presence of endogenous immunoglobulins.Alexa Fluor dyes are among the most trusted fluorescent dyes available today. Invitrogen™ Alexa Fluor 647 dye is a near-infrared-fluorescent dye with excitation ideally suited to the 647 nm laser line. For stable signal generation in imaging and flow cytometry, Alexa Fluor 647 dye is pH-insensitive over a wide molar range. Probes with high fluorescence quantum yield and high photostability allow detection of low-abundance biological structures with great sensitivity. Alexa Fluor 647 dye molecules can be attached to proteins at high molar ratios without significant self-quenching, enabling brighter conjugates and more sensitive detection. The degree of labeling for each conjugate is typically 2-8 fluorophore molecules per IgG molecule; the exact degree of labeling is indicated on the certificate of analysis for each product lot.Using conjugate solutions: Centrifuge the protein conjugate solution briefly in a microcentrifuge before use; add only the supernatant to the experiment. This step will help eliminate any protein aggregates that may have formed during storage, thereby reducing nonspecific background staining. Because staining protocols vary with application, the appropriate dilution of antibody should be determined empirically. For the fluorophore-labeled antibodies a final concentration of 1-10 µg/mL should be satisfactory for most immunohistochemistry and flow cytometry applications.Background/Target InformationWe offer an extensive line of Invitrogen™ secondary antibody conjugates with well-characterized specificity and labeled with a wide selection of premium fluorescent dyes, including Invitrogen™ Alexa Fluor™ fluorescent dyes. Fluorescent secondary antibody conjugates are useful in the detection, sorting, or purification of its specified target and ideal for fluorescence microscopy and confocal laser scanning microscopy, flow cytometry, and fluorescent western detection. The breadth of fluorescent markers we offer allows our reagents to be tailored to almost any fluorescent detection system.Secondary antibodies may be provided in three formats: whole IgG, divalent F(ab')2 fragments, and monovalent Fab fragments. Because of the high degree of conservation in the structure of many immunoglobulin domains, most class-specific secondary antibodies must be affinity-purified and cross-adsorbed to achieve minimal cross-reaction with other immunoglobulins.Our secondary antibody conjugates are most commonly prepared by immunizing the host animal with a pooled population of immunoglobulins from the target species and can be further purified and modified (e.g., immunoaffinity chromatography, antibody fragmentation, label conjugation, etc.) to generate highly specific reagents. In the first round of purification, whole immunoglobulins binding to the immunizing antibody are recovered and mainly consist of the ~150-kDa IgG class. Further purification, for example, with Protein A or G, removes all unwanted immunoglobulin classes except the affinity-purified antibodies that react with the target-specific immunoglobulin heavy and/or light chains.

数据

Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody (A-21246) in IFImmunofluorescence analysis of F (ab')2-Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor 647 (Product # A-21246) was performed using A-431 cells stained with EGFR (EP38Y) Rabbit Monoclonal Primary Antibody (Product # MA5-14485). The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, blocked with 1% BSA for 1 hour and labeled with 2 µg/mL of rabbit primary antibody for 3 hours at room temperature. F (ab')2-Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor 647 (Product # A-21246) was used at a concentration of 4 µg/mL in phosphate buffered saline containing 0.2 % BSA for 45 minutes at room temperature, for detection of EGFR in the membrane (Panel a: red). Nuclei (Panel b: blue) were stained with DAPI in SlowFade® Gold Antifade Mountant (Product # S36938). F-actin was stained with Alexa Fluor® 488 Phalloidin (Product # A12379, 1:300) (Panel c: green). Panel d represents the composite image. No nonspecific staining was observed with the secondary antibody alone (panel f), or with an isotype control (panel e). The images were captured at 60X magnification.

推荐产品：

二抗荧光二抗免疫组化一抗标签抗体

参考文献：

1. NatureCell diversity and network dynamics in photosensitive human brain organoids."A21246 was used in immunocytochemistry to compare the gene expression in individual cells isolated from human brain organoids"AuthorsQuadrato G,Nguyen T,Macosko EZ,Sherwood JL,Min Yang S,Berger DR,Maria N,Scholvin J,Goldman M,Kinney JP,Boyden ES,Lichtman JW,Williams ZM,McCarroll SA,Arlotta P2. OncotargetFAM65B controls the proliferation of transformed and primary T cells."A21246 was used in flow cytometry to study the role of FAM65B in cell proliferation"AuthorsFroehlich J,Versapuech M,Megrelis L,Largeteau Q,Meunier S,Tanchot C,Bismuth G,Delon J,Mangeney M3. Radiation researchDifferences in Radiation Dose Response between Small and Large Intestinal Crypts."A21246 was used in immunohistochemistry to assess the effects of middle- and low-dose radiation on intestinal stem cells"AuthorsOtsuka K,Suzuki K4. Proceedings of the National Academy of Sciences of the United States of AmericaDendritic compartmentalization of chloride cotransporters underlies directional responses of starburst amacrine cells in retina.AuthorsGavrikov KE,Nilson JE,Dmitriev AV,Zucker CL,Mangel SC

技术参数

产品应用 ICC;IF

© Bio-Rad伯乐代理官网-伯乐抗体-伯乐层析Aminex色谱柱是专业的授权总代理区域代理经销平台。
© 如需询价，请加客服QQ：1749072012 、客服微信：jinshanbio，或发送邮件到1749072012@qq.com
© 平台为生命科学研究相关领域提供一站式耗材试剂仪器解决方案和采购服务，数据资源基于CC协议。
© 本文地址：https://biorod.cn/thread-17279.htm