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WB: EGF-stimulated A431 cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with anti-Phosphotyrosine (2µg/ml). Prot...
Anti-Phosphotyrosine, clone PY20
Recommended Replacement for: 05-947 is a recommended replacement for MAB3080| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| Vrt | ELISA, IP, WB | Mouse | Purified | Monoclonal Antibody |
Description:
Anti-Phosphotyrosine, clone PY20
Replaces:
05-947 is a recommended replacement for MAB3080
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:
Phosphotyrosine conjugated to carrier protein.
Modifications:
Phosphorylation
Clone:
PY20
Isotype:
IgG2b
Background Information:
Tyrosine phosphorylation represents a major mechanism for the activation of signal transduction pathways in response to growth factors, hormones and cytokines. There are approximately 100 tyrosine kinases that are both receptor tyrsosine kinase receptors for various growth factors and cytokines. The extent and duration of tyrosine phosphorylation are tightly regulated by the 107 protein tyrosine phosphatases. Tyrosine phosphorylation occurs on residues on either other kinases (Ser/Thr or Tyr) or on substrates. In the case of kinases, this phosphorylation often causes the enzyme to change activation states and/or it creates binding sites for SH2 domain. Mutation or misregulation of protein tyrosine kinases is a cause or consequence of many disease states. It has been estimated that 20% of oncogenes so far described are tyrosine kinases. With such clear links between aberrant protein tyrosine kinase activity and disease, it is no surprise that much of the early effort in kinase drug discovery was focused on these enzymes.
Species Reactivity:
Vertebrates
Species Reactivity Note:
Most common vertebrate species tested. Reactivity with other species has not been confirmed.
Application Notes:
Immunoprecipitation:
Recommended by an independent laboratory to immunoprecipitate tyrosine phosphorylated proteins.
ELISA:
Recommended by an independent laboratory.
Recommended by an independent laboratory to immunoprecipitate tyrosine phosphorylated proteins.
ELISA:
Recommended by an independent laboratory.
Control:
EGF-stimulated A431 cell lysate.
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 by western blot with RIPA lysates EGF-treated human A431 carcinoma cells.
Western Blot Analysis:
0.5-2 μg/mL of this lot detected tyrosine-phosphorylated proteins in RIPA lysates EGFtreated human A431 carcinoma cells.
Western Blot Analysis:
0.5-2 μg/mL of this lot detected tyrosine-phosphorylated proteins in RIPA lysates EGFtreated human A431 carcinoma cells.
Purification Method:
Protein A purfied
Presentation:
Purified mouse monoclonal IgG2b in buffer containing 0.02 M PBS, 0.25 M NaCl, pH 7.6, with 0.1% sodium azide. Liquid at 2-8°C.
Storage Conditions:
Stable for 1 year at 2-8°C from date of recipt.
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.
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.
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:
- ELISA
- 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 PY20
Concentration:
1 mg/mL
Antibody Type:
Monoclonal Antibody
Qty/Pk:
1 mg
Format:
Purified
Host:
Mouse