F(ab')2-Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488

货号：A-11070

规格：500 µg

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 488

类型:	多抗	同型对照：	IgG
免疫原性：	Gamma Immunoglobins Heavy and Light chains	用法：	2.0 µg/mL(IHC);2.0 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 488 dye is a bright, green-fluorescent dye with excitation ideally suited to the 488 nm laser line. For stable signal generation in imaging and flow cytometry, Alexa Fluor 488 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 488 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 apropriate dilution of antibody should be determined empirically. For the fluoropore-labeled antibodies a final concentration of 1-10 µg/mL should be satisfactory for most immunohistochemistry and flow cytometry applications.

数据

Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody (A-11070) in IFImmunofluorescence analysis of F (ab')2-Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor 488 (Product # A-11070) was performed using HeLa cells stained with PARP Rabbit Polyclonal Primary Antibody (Product # PA5-16452). 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 488 (Product # A-11070) was used at a concentration of 2 µg/mL in phosphate buffered saline containing 0.2 % BSA for 45 minutes at room temperature, for detection of PARP in the nucleus (Panel a: green). Nuclei (Panel b: blue) were stained with DAPI in SlowFade® Gold Antifade Mountant (Product # S36938). F-actin was stained with Rhodamine Phalloidin (Product # R415, 1:300) (Panel c: red). 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.

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参考文献：

1.PloS one Death of Monocytes through Oxidative Burst of Macrophages and Neutrophils: Killing in Trans. 2.Nature Cell diversity and network dynamics in photosensitive human brain organoids. 3.PLoS genetics Retrotransposon activation contributes to neurodegeneration in a Drosophila TDP-43 model of ALS. 4.Scientific reports Mammalian TRAPPIII Complex positively modulates the recruitment of Sec13/31 onto COPII vesicles. 5.Scientific reports Progressive Muscle Cell Delivery as a Solution for Volumetric Muscle Defect Repair.

技术参数

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