HRP Protein A Antibody

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Other Names
Protein A
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HRPProteinA_WB_1_090616
Total cell lysates (15 µg protein) from HeLa cells were probed separately with purified anti-HSP70 antibody (clone W27), anti-HSP90 antibody (clone 3H3C27) and anti-β-actin antibody (clone Poly6221) followed by either goat anti-mouse-IgG secondary antibody conjugated to HRP, donkey anti-rabbit-IgG secondary antibody conjugated to HRP, or different concentrations of HRP Protein A.
  • HRPProteinA_WB_1_090616
    Total cell lysates (15 µg protein) from HeLa cells were probed separately with purified anti-HSP70 antibody (clone W27), anti-HSP90 antibody (clone 3H3C27) and anti-β-actin antibody (clone Poly6221) followed by either goat anti-mouse-IgG secondary antibody conjugated to HRP, donkey anti-rabbit-IgG secondary antibody conjugated to HRP, or different concentrations of HRP Protein A.
  • HRPProteinA_WB_2_090616
    IP followed by Western blot analysis. mCherry-fused protein was immunoprecipitated from 100 µg transfected 293E cell extract using 5 µg purified anti-mCherry antibody (clone 8C5.5). Immunoprecipitates and 10% of total lysates were probed with either goat anti-mouse-IgG secondary antibody conjugated to HRP or 0.25 µg/mL HRP Protein A. The arrows indicate IgG heavy chain (IgGHC) and IgG light chain (IgGLC).
Description

Protein A is a 42 kD surface protein originally found in the cell wall of the bacterium Staphylococcus aureus. It is widely used in biochemical research and industry because of its ability to bind immunoglobulins. It is composed of five homologous Ig-binding domains that fold into a three-helix bundle. Each domain is able to bind proteins from many mammalian species, most notably IgGs. It binds the heavy chain within the Fc region of most immunoglobulins. HRP Protein A is useful for detecting primary antibodies, e.g., in ELISA, ELISPOT, IHC, or Western blotting. For IP followed by Western blotting application, HRP protein A will avoid IgG heavy/light chain issue because it only recognizes the IgG in its native form, and the antibody used to IP the target is usually boiled with the immunoprecipitates and gets denatured.

Product Details
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689202 500 µl $40.00
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Technical data sheet

Product Details

Reactivity
Mouse, Rabbit
Formulation
This reagent is provided in 50% glycerol in aqueous buffered solutions with preservatives.
Preparation
Recombinant protein A was conjugated with HRP under optimal conditions.
Concentration
1.0 mg/ml
Storage & Handling
Upon receipt, the antibody solution should be stored undiluted at -20°C.
Application

WB - Quality tested

Recommended Usage

For Western blot analysis, the reagent should be titrated between 1:2000 - 1:8000 to determine optimal conditions.



Affinity of Protein A to Ig Types from Different Species:

Species

Immunoglobulin

Binding to Protein A

Mouse

IgG1

+

 

IgG2a

++++

 

IgG2b

+++

 

IgG3

++

 

IgM

-

Rat

IgG1

-

 

IgG2a

-

 

IgG2b

-

 

IgG2c

++

Human

Total IgG

++++

 

IgG1

++++

 

IgG2

++++

 

IgG3

-

 

IgG4

++++

 

IgM

-

 

IgA

-

 

IgE

-

Chicken

IgY

-

Goat

IgG

+/-

Hamster

IgG

+

Rabbit

IgG

++++

Sheep

IgG

+/-

Antigen Details

Structure
42 kD surface protein.
Function
Binds IgG.
Antigen References

1. Verwey WF. 1940. J. Exp. Med. 71:635.
2. Graille M, et al. 2000. Proc. Natl. Acad. Sci. USA 97:5399.

Gene ID
NA
UniProt
View information about Protein A on UniProt.org

Related FAQs

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Go To Top Version: 2    Revision Date: 09/12/2016

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