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WB: EGF-stimulated A431 cell lysate probed with anti-phosphotyrosine.
Anti-Phosphotyrosine
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H, R, M | IP, WB | Rabbit | Purified | Polyclonal Antibody |
Description:
Anti-Phosphotyrosine
Background Information:
The phosphorylation of specific tyrosine residues has been shown to be a primary mechanism of signal transduction during normal mitogenesis, cell cycle progression and oncogenic transformation, its role in other areas such as differentiation and gap junction communication, is a matter of active and ongoing research. Antibodies that specifically recognize phosphorylated tyrosine residues have proved to be invaluable to the study of tyrosine phosphorylated proteins and the biochemical pathways in which they function.
Specificity:
The immunoreactivity of the antibody is totally inhibited by the use of 100 mM phenyl phosphate, a phosphotyrosine analog. The phosphotyrosine antibody is purified by immunoaffinity chromatography using either a dual phospho-peptide gel or a BSA-phosphotyrosine gel. All of the phosphotyrosine immunoreactivity present in the antisera is immunoadsorbed whether the antibody is purified by either gel indicating that the antibody is not sequence-specific but specific for phosphotyrosine residues.
Key Applications:
- Immunoprecipitation
- Western Blotting
Application Notes:
Immunoprecipitation:
4 μg of a previous lot immunoprecipitated proteins containing phosphotyrosine residues from a human A431 RIPA lysate.
4 μg of a previous lot immunoprecipitated proteins containing phosphotyrosine residues from a human A431 RIPA lysate.
Species Reactivity:
- Human
- Rat
- Mouse
Species Reactivity Note:
Human, mouse and rat. Other species cross-reactivity is unknown.
Isotype:
IgG
Immunogen:
In order to produce broad spectrum polyclonal phosphotyrosine antibodies, rabbits were immunized with three phosphorylated immunogens: (1) phosphotyrosine covalently linked to KLH; (2) the c-Src carboxyl terminal regulatory phosphopeptide (TS-T-E-P-Q-pY-Q-P-G-E-N-L; Catalog # 12-218) covalently linked to KLH, and; (3) a phosphopeptide associated with high tyrosine kinase activity in human lymphocytes (R-R-L-I-E-D-A-E-pY-A-A-R-G; Catalog # 12-217) covalently linked to KLH. Both of the phosphopeptide haptens serve as strong substrates for tyrosine phosphatases and are part of the two colorimetric tyrosine phosphatase kits provided by Upstate, Inc. (Catalog # 17-125, 17-126).
Modifications:
Phosphorylation
Molecular Weight:
Varies
Control:
Included Positive Antigen Control:
Catalog # 12-302, EGF-stimulated A431 cell lysate. Add 2.5 μL of 2-mercaptoethanol/100 μL of lysate and boil for 5 minutes to reduce the preparation. Load 20 μg of reduced lysate per lane for minigels.
Catalog # 12-302, EGF-stimulated A431 cell lysate. Add 2.5 μL of 2-mercaptoethanol/100 μL of lysate and boil for 5 minutes to reduce the preparation. Load 20 μg of reduced lysate per lane for minigels.
Presentation:
Purified rabbit polyclonal in storage buffer containing 0.2 M Tris-glycine, Ph 7.2, 0.15 M NaCl, 1 mg/mL of BSA containing 0.05% sodium azide. Frozen at -20°C.
Storage Conditions:
Stable for 1 year at -20ºC from date of receipt.
Quality Assurance:
Evaluated by western blot on EGF-stimulated human A431 carcinoma cells.
Western Blot Analysis:
0.5-2 μg/mL of this antibody detected proteins containing phosphotyrosine residues in RIPA lysates from EGF-stimulated human A431 carcinoma cells.
Western Blot Analysis:
0.5-2 μg/mL of this antibody detected proteins containing phosphotyrosine residues in RIPA lysates from EGF-stimulated human A431 carcinoma cells.
Antibody Category:
Signaling
Antibody Sub-Category:
General Post-translation Modification
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Trade Name:
Upstate (Millipore)
Format Code:
Pur
Concentration:
181.81 µg/mL
Antibody Type:
Polyclonal Antibody
Format:
Purified
Host:
Rabbit
Purification Method:
Protein A Purified followed by immunoaffinity