STAT 5a,b, phosphorylated (Tyr694/699) (Signal Transducer and Activator of Transcription 5a,b)

Applications: \nSuitable for use in Western Blot, Peptide Inhibition and Peptide Dot Blot. Other applications not tested.\n\nRecommended Dilution:\nWestern Blot: 1:500-1:1000 detects phospho-STAT5A/B (Tyr694/699) in lysates from 3T3/NIH cells treated with PDGF. Non-treated and PDGF treated 3T3/NIH cells were resolved by electrophoresis, transferred to nitrocellulose and probed with antiphospho-STAT5A/B (Tyr694/699) (1:500 dilution). Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system.\n\nPeptide Inhibition: 1uM of the peptide containing the phospho-STAT5A/B (Tyr694/699) modification completely abolished detection of STAT5 by anti-phospho-STAT5A/B (Tyr694/699) in immunoblot analysis of 3T3/NIH cell lysates ( lane 2). 3T3/NIH cell lysates were resolved by electrophoresis, transferred to nitrocellulose and probed with antiphospho-STAT5A/B (Tyr694/699) (1:500 dilution) or antiphospho- STAT5A/B (Tyr694/699) pre-absorbed with 1uM peptides. Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system.\n\nPeptide Dot Blot: 1:500-1:2000 detected the peptide containing phospho-STAT5A/B (Tyr694/699).. A dilution series of peptides. 2ul of each dilution was spotted onto PVDF membranes and probed with anti-phospho-STAT5A/B (Tyr694/699) (1:1000 dilution). Peptides were visualized using a goat anti-rabbit secondary conjugated to HRP and a chemiluminescence detection system.\n\nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

S7972-14A

Size

100ul

Applications

WB

Hosts

Rabbit

Reactivities

Hum, Mouse, Rat, Bov

Form

Supplied as a liquid in 0.1M Tris-glycine, pH 7.4, 0.15M NaCl, 0.05% sodium azide before the addition of glycerol to 30%.

P Type

Pab

Purity

Purified by immunoaffinity chromatography.

Isotype

IgG

References

Gollob, J.A. et al., Blood 91: 1341-1354, 1998.

Additional Info

Recognizes phospho-STAT5A/B (Tyr694/699), Mr 95kD. An unknown protein was also\ndetected, Mr 200kD. Species Crossreactivity: Human, bovine, rat and mouse.