Goat Anti-Rabbit IgG (H+L) Secondary Antibody, Biotin Conjugated

This antibody is purified from antiserum by immunoaffinity chromatography which removes essentially all goat serum proteins, except the specific antibody for rabbit IgG. The antibody preparation is solid phase adsorbed with human serum proteins to ensure minimal cross reactivity in tissue or cell preparation

Product Info Summary

SKU: BA1003
Size: 0.5ml
Reactive Species: Rabbit
Host: Goat
Application: ELISA, IHC

Product Overview

Product Name Goat Anti-Rabbit IgG (H+L) Secondary Antibody, Biotin Conjugate
Synonyms Biotin-conjugated Goat Anti-Rabbit IgG; Goat Anti-Rabbit IgG Biotinylated Antibody; Biotinylated Goat Anti-Rabbit IgG Secondary Antibody; Goat Anti-Rabbit IgG Secondary Antibody, Biotin-labeled
Description Goat Anti-Rabbit IgG (H+L) Secondary Antibody, Biotin Conjugate, for indirect sensitive immunodetection and/or quantification of low-abundance target proteins through ELISA or IHC by using reporter-labeled biotin-binding signal amplification systems.
Reagent Type Biotin conjugated secondary antibody
Conjugate Biotin
Host Goat
Target Species Rabbit
Antibody Class IgG
Clonality Polyclonal
Immunogen Whole molecule rabbit IgG
Purification Immunoaffinity chromatography, solid-phase adsorbed with human serum proteins
Specificity Rabbit IgG specific; no cross-reactivity with human/bovine/mouse IgG
Form Supplied Liquid: concentrated buffered stock solution
Formulation 0.5 mg biotin-conjugated secondary antibody
0.01 M PBS (PH 7.4)
50% glycerol
Pack Size 0.5 ml
Concentration 1 mg/ml
Application ELISA, IHC
Storage At -20˚C for one year from date of receipt. Avoid repeated freezing and thawing.
Precautions FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR CLINICAL USE

Assay Information

Sample Type Human primary-antibody-probed: SDS-PAGE separated-, membrane-immobilized-proteins from cell/tissue lysates, formalin-fixed paraffin-embedded (FFPE) tissue sections on slides
Assay Type Immunoanalytical
Technique Indirect immunodetection of target protein via reporter-labeled biotin-binding detection systems
Assay Purpose Protein detection/quantification
Equipment Needed WB/Dot blot/ELISA/IHC instrumentation; Reporter signal detectors: X-ray film cassette; a charge-coupled device (CCD) imager; Spectrophotometer; fluorescent or electron microscope

Main Advantages

Specific High signal-to-noise ratio
High Signal Amplification Multiple secondary antibodies can bind to a single primary antibody; Multiple reporter molecules localize to a single biotin via avidin/streptavidin bridges
Fast Fewer processing steps - no need to add a substrate; Less optimization required compared to enzymatic detection; Generates strong signals in a relatively short time span; Fluorescence can be observed directly
Quantifiable Allows quantification of detected signal
Easy to Use Supplied in a workable liquid format
Flexible Biotin- (Strept)Avidin system can be coupled with various types of reporters (enzymes, fluorochromes, fluorophores, chromophores, etc.); One type of labeled secondary antibody can be used to recognize different types of primary antibodies of the target organism specific to a particular antigen
Compatible Biotin does not interfere with catalysis or antibody binding

Background

Most commonly, secondary antibodies are generated by immunizing the host animal with a pooled population of immunoglobulins from the target species. The host antiserum is then purified through immunoaffinity chromatography to remove all host serum proteins, except the specific antibody of interest. Purified secondary antibodies are further solid phase adsorbed with other species serum proteins to minimize cross-reactivity in tissue or cell preparations, and are then modified with antibody fragmentation, label conjugation, etc., to generate highly specific reagents. Secondary antibodies can be conjugated to a large number of labels, including enzymes, biotin, and fluorescent dyes/proteins. Here, the antibody provides the specificity to locate the protein of interest, and the label generates a detectable signal. The label of choice depends upon the experimental application.

Biotinylated antibodies are widely used in systems where signal amplification is desired. Often 15-20 biotin moieties are coupled to a single IgG secondary antibody. Biotin binds avidin, streptavidin, or neutravidin with a high degree of affinity and specificity. In immunoassays avidin/streptavidin-biotin binding is used as a bridge between antibodies and reporters like enzymes (HRP, AP), fluorophores, chromophores, etc. Both avidin and streptavidin are tetrameric proteins capable of binding 4 biotin groups to each molecule of avidin or streptavidin, thus amplifying the signal intensity and detection sensitivity by increasing the concentration of reporters at the antigenic site. Two main biotin-binding detection systems have been widely utilized: Avidin-Biotin Complex (ABC) and Labeled Streptavidin Biotin (LSAB) methods. In the ABC method free avidin (or streptavidin) is used as a bridge/link between the biotinylated antibody and а biotinylated reporter molecule, resulting in three reporter molecules coupled to the biotinylated antibody. The LSAB method employs a reporter-labeled streptavidin (avidin or neutravidin can alternatively be used) to detect the bound biotinylated-secondary antibody on the tissue section, blotting membrane or ELISA plate, improving the sensitivity of detection by 8-fold. The LSAB method is used when the avidin-biotin-enzyme complex in the ABC method becomes too large to penetrate the tissue.

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1 Customer Q&As for Goat Anti-Rabbit IgG (H+L) Secondary Antibody, Biotin Conjugated

Question

Would either BA1054 or BA1003 secondary antibody work for M00139 for flow cytometry?

Verified customer

Asked: 2019-11-25

Answer

We don't recommend the Goat Anti-Rabbit IgG (H+L) Secondary Antibody, HRP Conjugate (BA1054) and Goat Anti-Rabbit IgG (H+L) Secondary Antibody, Biotin Conjugate (BA1003) for M00139 for flow cytometry.

Boster Scientific Support

Answered: 2019-11-25

Size

Total: $95

SKU:BA1003

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