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05-947

Sigma-Aldrich

Anti-Phosphotyrosine Antibody, clone PY20

clone PY20, Upstate®, from mouse

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About This Item

UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41

biological source

mouse

Quality Level

100

antibody form

purified antibody

antibody product type

primary antibodies

clone

PY20, monoclonal

species reactivity (predicted by homology)

vertebrates (common species)

manufacturer/tradename

Upstate®

technique(s)

ELISA: suitable
immunoprecipitation (IP): suitable
western blot: suitable

isotype

IgG2b

shipped in

wet ice

target post-translational modification

phosphorylation (pTyr)

Gene Information

human ... PID1(55022)

General description

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.

Specificity

Reactivity with other species has not been confirmed.

Immunogen

Phosphotyrosine conjugated to carrier protein.

Application

Anti-Phosphotyrosine Antibody, clone PY20 is an antibody against Phosphotyrosine for use in ELISA, IP & WB.
Immunoprecipitation:
Recommended by an independent laboratory to immunoprecipitate tyrosine phosphorylated proteins.

ELISA:
Recommended by an independent laboratory.

Quality

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.

Target description

Dependent upon the molecular weight of the tyrosine phosphorylated protein being detected.

Linkage

Replaces: MAB3080

Physical form

0.1M Tris-Glycine, 0.15M NaCl, 0.05% Sodium Azide, pH 7.4. Liquid at 2-8°C.
Format: Purified

Storage and Stability

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.

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.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

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Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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J R Glenney et al.
Journal of immunological methods, 109(2), 277-285 (1988-05-09)
Phosphotyrosine coupled to KLH, BSA, and OVA was used for the production and screening of antibodies to phosphotyrosine. 800 hybridomas secreting antibodies that bound to phosphotyrosine were detected by ELISA. The most reactive 100 of these 800 were tested subsequently
Haoran Su et al.
PloS one, 7(2), e31636-e31636 (2012-02-24)
Leptin exerts its action by binding to and activating the long form of leptin receptors (LEPRb). LEPRb activates JAK2 that subsequently phosphorylates and activates STAT3. The JAK2/STAT3 pathway is required for leptin control of energy balance and body weight. Defects
Protein tyrosine phosphatases and signalling.
Stoker, Andrew W
The Journal of Endocrinology, 185, 19-33 (2005)
Inhibitors of protein tyrosine phosphatases: next-generation drugs?
Bialy, Laurent and Waldmann, Herbert
Angewandte Chemie (International ed. in English), 44, 3814-3839 (2005)
Katherine H Fisher et al.
Molecular biology of the cell, 27(3), 434-441 (2015-12-15)
Transmembrane receptors interact with extracellular ligands to transduce intracellular signaling cascades, modulate target gene expression, and regulate processes such as proliferation, apoptosis, differentiation, and homeostasis. As a consequence, aberrant signaling events often underlie human disease. Whereas the vertebrate JAK/STAT signaling
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