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Lysates from various cells with various treatments to stimulated varying tyrosine phosphorylations on multiple substrates were resolved in parallel by...
4G10® Platinum, Anti-Phosphotyrosine IHC(P) staining of various cancer tissue samples.
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Anti-Phosphotyrosine, 4G10® Platinum
| 交差性 | 主要アプリケーション | 免疫動物 | フォーマット | 抗体タイプ |
|---|---|---|---|---|
| A | WB, IC, ELISA, IH, IF, FC, IP, IPK | Mouse | Purified | Monoclonal Antibody |
Description:
4G10® Platinum, Anti-Phosphotyrosine (mouse monoclonal cocktail IgG2b)
Background Information:
The development of the anti-phosphotyrosine, clone 4G10® in 1989 was a monumental discovery for researchers. 4G10® was the first and is the best single monoclonal antibody for the detection tyrosine phosphorylation. 4G10® is well known for its sensitivity and its ability to detect multiple tyrosine phosphorylations on numerous substrates. It has been validated by thousands of scientific and medical researchers in virtually every application and tyrosine target over the past 2 decades. To improve on something that hundred have tried and no one has succeeded, we pooled 4G10® with the next most highly regarded anti-phosphotyrosine, clone PY20 to make 4G10® Platinum. PY20 itself is a very poor substitute for 4G10®, but its additive effect allow for a greater level of detection on more substrates that even 4G10® alone was not capable of.
Specificity:
Tyrosine-phosphorylated proteins from all species
Key Applications:
- Western Blotting
- Immunocytochemistry
- ELISA
- Immunohistochemistry
- Immunofluorescence
- Flow Cytometry
- Immunoprecipitation
- IP-Kinase Assay
Application Notes:
Immunoprecipitation: 2-4 μL of a previous lot can immunoprecipitate quantitatively the phosphotyrosine containing proteins in the lysate of a confluent culture (10cm dish) of cells expressing an activated tyrosine kinase. To preserve phosphotyrosine, add 0.2 mM sodium orthovanadate to the lysis buffer.
Species Reactivity:
All
Isotype:
IgG2b
Immunogen:
4G10® used Phosphotyramine coupled to KLH and PY20 used phosphotyrosine conjugated to carrier protein
Modifications:
Phosphorylation
Molecular Weight:
Dependent upon the molecular weight of the tyrosine phosphorylated protein being detected.
Control:
Suggested Positive Antigen Control: Cat. # 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 -20ºC. The lysate is stable for 6 months at -20ºC. Before use, add 2.5 μL of 2-mercaptoethanol per 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:
Proprietary mixture of protein G purified mouse monoclonals 4G10® (IgG2b) and PY20 (IgG2b) in 0.1 M Tris-glycine, Ph 7.4, 0.15 M NaCl, 0.05% sodium azide.
Liquid at 2-8°C.
Liquid at 2-8°C.
Storage Conditions:
Stable for 1 year at from date of receipt.
Quality Assurance:
Evaluated by Western Blot on a modified RIPA lysate from EGF-treated human A431 carcinoma cells.
Western Blot Analysis: 1:1,000-1:2,000 of this antibody detected tyrosinephosphorylated proteins in a modified RIPA lysate from EGF-treated human A431 carcinoma cells.
Western Blot Analysis: 1:1,000-1:2,000 of this antibody detected tyrosinephosphorylated proteins in a modified RIPA lysate from EGF-treated human A431 carcinoma cells.
Antibody Category:
Signaling
Antibody Sub-Category:
General Post-translation Modification
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.
Trade Name:
Upstate (Millipore)
Antibody Type:
Monoclonal Antibody
Format:
Purified
Host:
Mouse
Purification Method:
Protein G-Sepharose chromatography