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|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)|
|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)
|Pack Size||0.5 ml|
*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|
|Sample Type||SDS-PAGE separated-, membrane-immobilized-, mouse primary-antibody-probed proteins from cell/tissue lysates|
|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
|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|
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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