05-321X
Anti-Phosphotyrosine Antibody, clone 4G10®
clone 4G10®, Upstate®, from mouse
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UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41
biological source
mouse
Quality Level
antibody form
purified immunoglobulin
antibody product type
primary antibodies
clone
4G10®, monoclonal
species reactivity
vertebrates
manufacturer/tradename
Upstate®
technique(s)
immunocytochemistry: suitable
immunohistochemistry: suitable
immunoprecipitation (IP): suitable
western blot: suitable
isotype
IgG2bκ
shipped in
wet ice
target post-translational modification
phosphorylation (pTyr)
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.
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
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.
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.
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.
Research Category
Signaling
Signaling
Research Sub Category
General Post-translation Modification
General Post-translation Modification
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).
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.
Physical form
Format: Purified
Protein G-Sepharose Chromatography
Purified mouse monoclonal IgG2bκ in buffer containing 0.1 M Tris-glycine, pH 7.4, 0.15 M NaCl, 0.05% sodium azide. Liquid at 2-8ºC.
IgG2bκ mouse monoclonal antibody produced in vitro by mouse-mouse hybridoma 4G10 (FOX-NY [NS-1 derivative] myeloma x spleen cells).
IgG2bκ mouse monoclonal 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.
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
Positive Antigen Control: Catalog #12-302, EGF-stimulated A431 cell lysate. Add 2.5µL of 2-mercaptoethanol/100µL of lysate and boil for 5 minutes to reduce the preparation. Load 20µg of reduced lysate per lane for minigels.
Positive Antigen Control: Catalog #12-302, EGF-stimulated A431 cell lysate. Add 2.5µL of 2-mercaptoethanol/100µL of lysate and boil for 5 minutes to reduce the preparation. Load 20µg of reduced lysate per lane for minigels.
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.
wgk_germany
nwg
flash_point_f
Not applicable
flash_point_c
Not applicable
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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