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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| Vrt | ICC, IHC, IP, WB | M | Purified | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | 05-321X |
| Brand Family | Upstate |
| Trade Name |
|
| 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 |
|
| Presentation | 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). |
| Quality Level | MQ100 |
| 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 |
|
| 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 |
|
| Antibody Type | Monoclonal Antibody |
| Modifications |
|
| 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 |
|
| 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. |
| Packaging Information | |
|---|---|
| Material Size | 50 µg |