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05-321

Sigma-Aldrich

Anti-Phosphotyrosine Antibody, clone 4G10®

clone 4G10®, Upstate®, from mouse

eCl@ss:
32160702
NACRES:
NA.41

Quality Level

biological source

mouse

antibody form

purified antibody

antibody product type

primary antibodies

clone

4G10®, monoclonal

species reactivity (predicted by homology)

all

manufacturer/tradename

Upstate®

technique(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.

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificate of Analysis

Certificate of Origin

Genome-scale analysis reveals Sst2 as the principal regulator of mating pheromone signaling in the yeast Saccharomyces cerevisiae.
Chasse, SA; Flanary, P; Parnell, SC; Hao, N; Cha, JY; Siderovski, DP; Dohlman, HG
Eukaryotic Cell null
Expansion of natural killer cells with lytic activity against autologous blasts from adult and pediatric acute lymphoid leukemia patients in complete hematologic remission.
Giovanni F Torelli, Anna Guarini, Roberta Maggio, Cecilia Alfieri, Antonella Vitale, Robin Foa
Haematologica null
Tyrosine phosphorylation regulates nuclear translocation of PKCdelta.
Humphries, MJ; Ohm, AM; Schaack, J; Adwan, TS; Reyland, ME
Oncogene null
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

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

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