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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 | Mouse | Purified | Monoclonal Antibody |
Description:
4G10® Platinum, Anti-Phosphotyrosine (mouse monoclonal cocktail IgG2b)
Promotional Text:
FREE Positive Control Included:
EGF-stimulated A431 cell lysate (cat. # 12-302)
EGF-stimulated A431 cell lysate (cat. # 12-302)
Trade Name:
Upstate (Millipore)
Specificity:
Tyrosine-phosphorylated proteins from all species
Molecular Weight:
Dependent upon the molecular weight of the tyrosine phosphorylated protein being detected.
Immunogen:
4G10® used Phosphotyramine coupled to KLH and PY20 used phosphotyrosine conjugated to carrier protein
Modifications:
Phosphorylation
Isotype:
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.
Species Reactivity:
All
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.
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.
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.
Purification Method:
Protein G-Sepharose™ chromatography
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.
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.
Key Applications:
- Western Blotting
- Immunocytochemistry
- ELISA
- Immunohistochemistry
- Immunofluorescence
- Flow Cytometry
- Immunoprecipitation
Brand Family:
Upstate
Protein/Isoform Description:
Protein Tyrosine phosphorylation was first described in 1979. The development of the Anti-Phosphotyrosine, clone 4G10 in 1989 was a monumental discovery for researchers. 4G10 is the first and best single monoclonal antibody for the detection of tyrosine phosphorylation. It is the most highly cited (75% of related scientific journal articles over the last three years) and validated phosphotyrosine antibody available. It consistently produces high quality, reproducible data, and has been validated by thousands of scientific and medical researchers in virtually every application and tyrosine target over the past 18 years. In addition to its unparalleled characterization and applicability, 4G10 is well known as one of the most sensitive anti-phosphotyrosines. Now Millipore is excited to announce that we have taken the sensitivity for phosphotyrosine detection to the next level. To improve on something that hundreds have tried and none have 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 allows for a greater level of detection on more substrates that even 4G10 alone was capable of.
Product Name:
Anti-Phosphotyrosine, 4G10® Platinum
Antibody Type:
Monoclonal Antibody
Qty/Pk:
100 µL
Format:
Purified
Host:
Mouse