Product Pathways - Jak/Stat Pathway
Stat1 Blocking Peptide (9175 Specific) #1079
Description
This peptide is used to block Stat1 (42H3) Rabbit mAb #9175 reactivity.
Quality Control
The quality of the peptide was evaluated by reversed-phase HPLC and by mass spectrometry. The peptide blocks Stat1 (42H3) Rabbit mAb #9175 in immunohistochemistry and Western blot.
Western Blotting
Western blot analysis of extracts from serum starved HeLa cells, untreated or IFN-alpha-treated (100 ng/ml for 5 mintues), using Stat1 (42H3) Rabbit mAb #9175 preincubated with an unrelated control peptide (left) or Stat1 Blocking Peptide (#9175 specific) #1079 (right).
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using Stat1 (42H3) Rabbit mAb (Human Specific) #9175 in the presence of control peptide (left) or Stat1 Blocking Peptide (9175 Specific) (right).
Applications
Use as a blocking reagent to evaluate the specificity of antibody reactivity in immunohistochemistry and Western blot protocols.
Directions for Use
For immunohistochemistry, add twice the volume of peptide as volume of antibody used in 100 µl total volume. Incubate for a minimum of 30 minutes prior to adding the entire volume to the slide. Recommended antibody dilutions can be found on the relevant product data sheet.
For Western immunoblotting, add 10 µl of antibody and 10 µl of blocking peptide to 10 ml of antibody dilution buffer, and incubate at room
temperature for 30 minutes before allowing to react with the blot.
Background
The Stat1 transcription factor is activated in response to a large number of ligands (1) and is essential for responsiveness to IFN-α and IFN-γ (2,3). Phosphorylation of Stat1 at Tyr701 induces Stat1 dimerization, nuclear translocation and DNA binding (4). Stat1 protein exists as a pair of isoforms, Stat1α (91 kDa) and the splice variant Stat1β (84 kDa). In most cells, both isoforms are activated by IFN-α, but only Stat1α is activated by IFN-γ. The inappropriate activation of Stat1 occurs in many tumors (5). In addition to tyrosine phosphorylation, Stat1 is also phosphorylated at Ser727 through a p38 mitogen-activated protein kinase (MAPK)-dependent pathway in response to IFN-α and other cellular stresses (6). Serine phosphorylation may be required for the maximal induction of Stat1-mediated gene activation.
- Heim, M.H. (1999) J. Recept. Signal. Transduct. Res. 19, 75-120.
- Durbin, J.E. et al. (1996) Cell 84, 443-450.
- Meraz, M.A. et al. (1996) Cell 84, 431-442.
- Ihle, J.N. et al. (1994) Trends Biochem. Sci. 19, 222-227.
- Frank, D.A. (1999) Mol. Med. 5, 432-456.
- Wen, Z. et al. (1995) Cell 82, 241-250.
Application References
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