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Sigma-Aldrich

Anti-Phosphotyrosine Antibody, clone 4G10®, Biotin Conjugate

clone 4G10®, Upstate®, from mouse

Synonym(s):

4G10 Biotin Antibody, Biotinylated Anti-Phosphotyrosine, Clone 4G10 Anti-Phosphotyrosine

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About This Item

UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41

biological source

mouse

Quality Level

conjugate

biotin conjugate

antibody form

purified antibody

antibody product type

primary antibodies

clone

4G10®, monoclonal

species reactivity

human

species reactivity (predicted by homology)

all

manufacturer/tradename

Upstate®

technique(s)

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

isotype

IgG2bκ

shipped in

wet ice

target post-translational modification

phosphorylation (pTyr)

Gene Information

human ... PID1(55022)

General description

Anti-phosphotyrosine monoclonal antibody, clone 4G10 (cat.# 05-321), cross-linked to biotin.
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-phospho-tyrosine 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 response to various treatments.

Description
Anti-phosphotyrosine monoclonal antibody, clone 4G10 (cat. # 05-321), cross-linked to biotin.

Specificity

Expected to cross-react with rat and mouse.
Recognizes tyrosine-phosphorylated proteins from all species.

Immunogen

Phosphotyramine-KLH.

Application

Anti-Phosphotyrosine Antibody, clone 4G10, Biotin Conjugate detects tyrosine phosphorylated proteins in all species. This unique monoclonal antibody is published in peer reviewed journals and validated for use in IC, IP, WB.
Immunoprecipitation:
4 μg of a previous lot, used in conjunction with Streptavidin, agarose conjugate (Catalog # 16-126), immunoprecipitated phosphotyrosine containing proteins from a lysate of EGFstimulated A431 cells.
Note: To preserve phosphotyrosine, add 0.2 mM sodium orthovanadate to the lysis buffer.

Immunocytochemistry:
5-10 μg/mL of a previous lot gave positive immunostaining of EGF-stimulated A431 cells that had been fixed with ethanol:acetic acid [1:1].

Application Notes
For use in applications in which a biotin conjugate is advantageous, such as WB and IC.
Research Category
Signaling
Research Sub Category
General Post-translation Modification

Quality

Routinely evaluated by Western Blot on EGF treated A431 lysates.

Western Blot Analysis:
1:500 dilution of this lot detected Tyrosine phosphorylated proteins on 10 μg of EGF treated A431 lysates.

Target description

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

Physical form

PBS containing 0.05% sodium azide.
Protein G Purified

Storage and Stability

9 months at 2-8°C from date of shipment. NOTE: DO NOT FREEZE. For maximum recovery of product, centrifuge the original vial prior to removing the cap.

Analysis Note

Control
Pervanadate-treated human A431 cell extracts or EGF-treated human A431 cells

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.

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Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 2

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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H Nishizumi et al.
The Journal of experimental medicine, 187(8), 1343-1348 (1998-05-23)
B cells from young lyn-/- mice are hyperresponsive to anti-IgM-induced proliferation, suggesting involvement of Lyn in negative regulation of B cell antigen receptor (BCR)-mediated signaling. Here we show that tyrosine phosphorylation of FcgammaRIIB and CD22 coreceptors, which are important for
Sandra Odom et al.
The Journal of experimental medicine, 199(11), 1491-1502 (2004-06-03)
A role for Lyn kinase as a positive regulator of immunoglobulin (Ig)E-dependent allergy has long been accepted. Contrary to this belief, Lyn kinase was found to have an important role as a negative regulator of the allergic response. This became
Eivind F Finne et al.
The Biochemical journal, 379(Pt 1), 39-46 (2003-12-25)
Ephrins are ligands for the Eph receptor tyrosine kinases, which play important roles in patterning nervous and vascular systems. Ephrin-A1 is a glycosylphosphatidylinositol-anchored ligand that binds to the EphA receptor tyrosine kinases. In the present study, we have identified a
Yasuko Furumoto et al.
The Journal of biological chemistry, 279(47), 49177-49187 (2004-09-10)
The high affinity IgE Fc receptor (FcepsilonRI) beta chain functions as a signal amplifier and has been linked to atopy, asthma, and allergy. Herein, we report on a previously unrecognized negative regulatory role for the nonconventional beta chain immunoreceptor tyrosine-based
Imran H Khan et al.
Molecular & cellular proteomics : MCP, 5(4), 758-768 (2005-12-22)
Phosphorylation analysis of signaling proteins is key for examining intracellular signaling pathways. Conventional biochemical approaches, e.g. immunoprecipitation, Western blot, and ELISA, have played a major role in elucidation of individual signaling events. However, these methods are laborious, time-consuming, and difficult

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