HRP Conjugated AffiniPure Goat Anti-Rabbit IgG (H+L)

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 preparations.

Product Info Summary

SKU BA1054
Pack size 0.5ml
Applications WB, ELISA

Product Overview

Product Name HRP Conjugated AffiniPure Goat Anti-Rabbit IgG (H+L)
Synonyms HRP-conjugated goat anti-rabbit IgG
Description Goat Anti-Rabbit IgG (H+L) Secondary Antibody, HRP Conjugate, for the indirect sensitive immunodetection and/or quantification of target proteins through WB, or ELISA by assaying an HRP-catalyzed reaction product in the vicinity of the antigen-primary antibody-secondary antibody-HRP complex.
Reagent Type Secondary antibody, reporter enzyme labeled
Label HRP (Horseradish Peroxidase)
Host Goat
Target Species Rabbit
Antibody Class IgG
Clonality Polyclonal
Immunogen Rabbit IgG (whole molecular)
Purification The antibody was purified from antisera by immunoaffinity chromatography.
Specificity This HRP conjugated antibody is specific for rabbit IgG and shows no cross-reactivity with mouse/bovine IgG.
Form Supplied Concentrated, Liquid
Formulation 0.5 mg of HRP conjugated specific antibody
0.01 M PBS (PH 7.4)
50% glycerol
Pack Size 0.5 ml
Concentration 1mg/ml
Application ELISA*, WB*
*Our Boster Guarantee covers the use of this product in the above marked tested applications.
Storage At 4 ˚C for 12 months from the date of receipt.
Precautions FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR CLINICAL USE

Assay Information

Sample Type SDS-PAGE separated-, membrane-immobilized-, mouse primary-antibody-probed proteins from cell/tissue lysates
Assay Type Immunoassay
Assay Purpose Protein detection/quantification
Technique Immunodetection of target antibody with HRP reporter enzyme
Equipment Needed WB/ELISA instrumentation; X-ray film cassette or a charge-coupled device (CCD) imager; Spectrophotometer
Compatibility with Reagents Incompatible with sodium azide and metals
incompatible with high phosphate concentrations

Main Advantages

Specific High signal-to-noise ratio
Sensitive Detects low-abundant targets due to an optimal number of HRP molecules per antibody
High Signal Amplification Multiple secondary antibodies can bind to a single primary antibody;Secondary antibodies Fc regions provide further binding locations for biotin, or enable the use of ABC and SABC
Fast Generates strong signals in a relatively short time span
Quantifieable Allows quantification of detected signal
Easy to Use Supplied in a workable liquid format
Flexible HRP: compatible with chromogenic, fluorogenic and chemiluminescent substrates;
Convenient HRP’s small size: no interference with the primary/secondary antibody interaction; no steric hindrance to antibody/antigen complexes

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, and are then modified with antibody fragmentation, label conjugation, etc. 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.

Horseradish peroxidase (HRP) is extensively used for labeling secondary antibodies in ELISA, western blot, dot blot and immunohistochemistry. The HRP enzyme is made visible using a substrate that, when oxidized by HRP in the presence of hydrogen peroxide as an oxidizing agent, yields a characteristic change that is detectable by specific detection methods. The substrates commonly used with HRP fall into different categories including chromogenic, fluorogenic, and chemiluminescent substrates depending on whether they produce a colored, fluorimetric or luminescent derivative respectively. The intensity of the signal is proportional to peroxidase activity and is a measure of the number of enzyme molecules reacting, hence of the amount of recognized primary antibodies, and thus of the amount of target antigen.

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BA1054-2
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