05-321

Sigma-Aldrich

Anti-Phosphotyrosine Antibody, clone 4G10®

clone 4G10®, Upstate®, from mouse

eCl@ss:
32160702
NACRES:
NA.41
Pricing and availability is not currently available.

Quality Level

biological source

mouse

antibody product type

primary antibodies

clone

4G10®, monoclonal

species reactivity (predicted by homology)

all

mfr. no.

Upstate®

application(s)

immunocytochemistry: suitable
immunohistochemistry: suitable
immunoprecipitation (IP): suitable
western blot: suitable

isotype

IgG2bκ

shipped in

wet ice

General description

Some of the tyrosine residues can be tagged with a phosphate group (phosphorylated) by protein kinases. (In its phosphorylated state, it is referred to as phosphotyrosine.). Tyrosine phosphorylation is considered as one of the key steps in signal transduction and regulation of enzymatic activity.
The advent of anti-phosphotyrosine antibodies is one of significant events in signal transduction research. Before the availability of anti-phosphotyrosine antibodies, tyrosyl phospyhorylation of proteins and enzymes was investigated through hazardous and time-consuming radioactive experiments. Anti-phosphotyrosine antibodies are commonly used in western blots after the targeted proteins have been immunoprecipitated to measure the tyrosyl phosphorylation of the proteins. Anti-phosphotyrosine antibodies are also directly used on cell lysate to examine the overall change of tyrosine phosphorylation level in reponse to various treatments.

Specificity

This antibody recognizes tyrosine-phosphorylated proteins from all species.

Immunogen

Phosphotyramine-KLH

Application

Research Sub Category
General Post-translation Modification
Anti-Phosphotyrosine Antibody, clone 4G10 detects tyrosine phosphorylated proteins in all species. This unique monoclonal antibody is validated for use in IC, IH, IP, WB and is backed by hundreds of publications
Research Category
Signaling
Immunoprecipitation:
2-4 μg of a previous lot immunoprecipitated quantitatively the phosphotyrosine containing proteins in the lysate of a confluent culture (10 cm dish) of cells expressing an activated tyrosine kinase. To preserve phosphotyrosine, add: 0.2 mM sodium orthovanadate to the lysis buffer.

Quality

Routinely evaluated on EGF-treated human A431 carcinoma cells.

Western Blot Analysis:
0.5-2 μg/mL of this lot detected tyrosine-phosphorylated proteins in a modified RIPA lysate from EGF-treated human A431 carcinoma cells (Cohen, B., 1990; , Druker, B. J., 1989; Kanakura, Y., 1991).

Target description

Dependent upon the molecular weight of the tyrosine phosphorylated protein being detected.

Linkage

Replaces: MAB3090

Physical form

Protein G-Sepharose Chromatography
Format: Purified
0.1M Tris-Glycine, 0.15M NaCl, 0.05% Sodium Azide, pH 7.4. Liquid at 2-8°C.
IgG2bκ mouse monocolonal antibody produced in vitro by mouse-mouse hybridoma 4G10 (FOX-NY NS-1 derivative myeloma x spleen cells).

Storage and Stability

Stable for 1 year at 2-8°C from date of receipt.

NOTE: DO NOT FREEZE.
For maximum recovery of the product, centrifuge the original vial prior to removing the cap. If the product has accidentally been frozen and thawed, spin it at 13,000 x g for 10 minutes at 2-8ºC. Save the supernatant for application.

Analysis Note

Control
Untreated A-431 (negative control) and EGF treated A-431 (positive control) whole cell lysates.
Included Positive Antigen Control: Catalog # 12-302, EGF-stimulated A431 cell lysate is provided as a free positive antigen control for western immunoblotting. Aliquot as desired, refreeze immediately, and store at -20°C. The lysate is stable for 6 months at -20°C. Before use, 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 immunoblot analysis.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Legal Information

4G10 is a registered trademark of Upstate Group, Inc.
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Disclaimer

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.
Human neutrophils as a source of nociceptin: a novel link between pain and inflammation.
Fiset, ME; Gilbert, C; Poubelle, PE; Pouliot, M
Biochemistry null
Hyaluronidase 2 negatively regulates RON receptor tyrosine kinase and mediates transformation of epithelial cells by jaagsiekte sheep retrovirus.
Danilkovitch-Miagkova, A; Duh, FM; Kuzmin, I; Angeloni, D; Liu, SL; Miller, AD; Lerman, MI
Proceedings of the National Academy of Sciences of the USA null
Dmitry E Shvartsman et al.
The Journal of cell biology, 178(4), 675-686 (2007-08-19)
Src functions depend on its association with the plasma membrane and with specific membrane-associated assemblies. Many aspects of these interactions are unclear. We investigated the functions of kinase, SH2, and SH3 domains in Src membrane interactions. We used FRAP beam-size...
Neoplastic transformation by the gep oncogene, Galpha12, involves signaling by STAT3.
Kumar, RN; Shore, SK; Dhanasekaran, N
Oncogene null
Neil S Magoski et al.
The Journal of neuroscience : the official journal of the Society for Neuroscience, 25(35), 8037-8047 (2005-09-02)
Although ion channels are regulated by protein kinases, it has yet to be established whether the behavioral state of an animal may dictate whether or not modulation by a kinase can occur. Here, we describe behaviorally relevant changes in the...

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