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millipore Phosphotyrosine Antibody, clone 4G10抗體

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millipore Phosphotyrosine Antibody, clone 4G10抗體

05-321 | Anti-Phosphotyrosine Antibody, clone 4G10

Replacement Information 05-321 is a recommended replacement for MAB3090

millipore Phosphotyrosine Antibody, clone 4G10抗體

Species Reactivity	Key Applications	Host	Format	Antibody Type
A	ICC, WB, IP, IHC	M	Purified	Monoclonal Antibody

Description
Catalogue Number	05-321
Replaces	MAB3090
Brand Family	Upstate
Trade Name	Upstate
Description	Anti-Phosphotyrosine Antibody, clone 4G10?
Background Information	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.

Product Information
Format	Purified
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 immediay, 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.
Presentation	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).

Applications
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
Key Applications	Immunocytochemistry Western Blotting Immunoprecipitation Immunohistochemistry
Application Notes	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.

Biological Information
Immunogen	Phosphotyramine-KLH
Clone	4G10?
Concentration	Please refer to the Certificate of Analysis for the lot-specific concentration.
Host	Mouse
Specificity	This antibody recognizes tyrosine-phosphorylated proteins from all species.
Isotype	IgG2bκ
Species Reactivity	All
Antibody Type	Monoclonal Antibody
Modifications	Phosphorylation
Purification Method	Protein G-Sepharose? Chromatography
Molecular Weight	Dependent upon the molecular weight of the tyrosine phosphorylated protein being detected.

Product Usage Statements
Quality Assurance	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).
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.

Storage and Shipping Information
Storage Conditions	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.