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- Regulatory Status
- RUO
- Other Names
- Protein A
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- Product Citations
- publications
-
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. -
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).
| Cat # | Size | Price | Quantity Check Availability | Save | ||
|---|---|---|---|---|---|---|
| 689202 | 500 µL | $47 | ||||
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 DetailsProduct Details
- Verified 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, and protected from prolonged exposure to light.
- 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
+/-
- Product Citations
-
Antigen Details
- Structure
- 42 kD surface protein.
- Function
- Binds IgG.
- Biology Area
- Cell Biology
- 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
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