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Ordering Information
- : 05-321
- : 100 µg
- : 1 mg/mL
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WB: 20 micrograms of EGF-stimulated A431 cell lysate was probed with 1µg/ml of Anti-Phosphotyrosine, clone 4G10®.
Anti-Phosphotyrosine, clone 4G10®
Recommended Replacement for: 05-321 is a recommended replacement for MAB3090| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| A | IC, IH, IP, WB | Mouse | Purified | Monoclonal Antibody |
Description:
Anti-Phosphotyrosine, clone 4G10®
Replaces:
05-321 is a recommended replacement for MAB3090
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:
This antibody recognizes tyrosine-phosphorylated proteins from all species.
Molecular Weight:
Dependent upon the molecular weight of the tyrosine phosphorylated protein being detected.
Immunogen:
Phosphotyramine-KLH
Modifications:
Phosphorylation
Clone:
4G10®
Isotype:
IgG2bκ
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.
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.
Species Reactivity:
All
Species Reactivity Note:
All species
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.
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.
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 immediately, 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.
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 immediately, 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.
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).
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).
Purification Method:
Protein G-Sepharose™ chromatography
Presentation:
Purified mouse monoclonal IgG2bκ in buffer containing 0.02 M phosphate buffer, pH 7.6, 0.25 M NaCl, 0.1% sodium azide. Liquid at 2-8ºC.
IgG2bκ mouse monocolonal antibody produced in vitro by mouse-mouse hybridoma 4G10® (FOX-NY [NS-1 derivative] myeloma x spleen cells).
IgG2bκ mouse monocolonal antibody produced in vitro by mouse-mouse hybridoma 4G10® (FOX-NY [NS-1 derivative] myeloma x spleen cells).
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.
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.
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:
- Immunocytochemistry
- Immunohistochemistry
- Immunoprecipitation
- Western Blotting
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, clone 4G10®
Concentration:
1 mg/mL
Antibody Type:
Monoclonal Antibody
Qty/Pk:
100 µg
Format:
Purified
Host:
Mouse