Description
General description
Dependent upon the molecular weight of the tyrosine phosphorylated protein being detected.
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.
Immunogen
4G10 used Phosphotyramine coupled to KLH and PY20 used phosphotyrosine conjugated to carrier protein.
Application
Anti-Phosphotyrosine Antibody, 4G10 Platinum is an antibody against Phosphotyrosine for use in WB, IF, FC, IH, IP, IC, ELISA.
Research Category
Signaling
Research Sub Category
General Post-translation Modification
Biochem/physiol Actions
Tyrosine-phosphorylated proteins from all species
Physical form
A proprietary mixture of protein G purified mouse monoclonals 4G10 (IgG2bκ) and PY20 (IgG2b) in 0.1 M Tris-glycine, pH 7.4, 0.15 M NaCl, 0.05% sodium azide.
Format: Purified
Protein G-Sepharose Chromatography
Preparation Note
2 years at 2-8°C from date of shipment.
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 2-8°C. Save the supernatant for application.
Analysis Note
Control
Positive Antigen Control: Catalog #12-302, EGF-stimulated A431 cell lysate. Add 2.5µL of 2-mercaptoethanol/100µL of lysate and boil for 5 minutes to reduce the preparation. Load 20µg of reduced lysate per lane for minigels.
Routinely evaluated by immunoblot on a modified RIPA lysate from EGF-treated human A431 carcinoma cells
Legal Information
4G10 is a registered trademark of Upstate Group, Inc.
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
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.