Product Pathways - Jak/Stat Pathway
Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb #9167
PhosphoSitePlus® protein, site, and accession data: STAT1
| Applications | Reactivity | Sensitivity | MW (kDa) | Isotype |
|---|---|---|---|---|
| W IP IHC-P IHC-F IF-IC F ChIP | H M | Endogenous | 84, 91 | Rabbit IgG |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IHC-P=Immunohistochemistry (Paraffin)
IHC-F=Immunohistochemistry (Frozen)
IF-IC=Immunofluorescence (Immunocytochemistry)
F=Flow Cytometry
ChIP=Chromatin IP
Reactivity Key:
H=Human
M=Mouse
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
- 9167:
- ChIP Agarose, ChIP Magnetic, Flow, IHC / Frozen, IHC / Paraffin, Immunofluorescence*, Immunoprecipitation, Western Blotting
* Product-specific protocol.
Specificity / Sensitivity
Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb detects endogenous levels of Stat1 only when phosphorylated at tyrosine 701. The antibody detects phosphorylated tyrosine 701 of p91 Stat1 and also the p84 splice variant. It does not cross-react with the corresponding phospho-tyrosines of other Stat proteins.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr701 of human Stat1.
Western Blotting
Western blot analysis of extracts from HeLa cells untreated or treated with interferon-α (IFN-α), using Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb (upper) or Stat1 Antibody (#9172) (lower).
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded Non-Hodgkin's lymphoma control (left) or λ phosphatase treated (right), using Phospho-Stat1 (tyr701) (58D6) Rabbit mAb.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded Non-Hodgkin’s lymphoma using Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb #9167 in the presence of control peptide (left) or Phospho-Stat1 (Tyr701) Blocking Peptide (right).
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded stomach (chronic gastritis), using Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb.
IHC-P (paraffin)
Immunohistochemical analysis using Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb on SignalSlide® Phospho-Stat1/3/5 IHC Controls #8105 (paraffin-embedded HeLa cell pellets, untreated (left), treated with Human Interferon-α1 (hIFN-α1) #8927 (middle), or treated with Human Epidermal Growth Factor (hEGF) #8916 (right).
IHC-F (frozen)
Immunohistochemical analysis of frozen SKOV-3 xenograft using Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb.
Flow Cytometry
Flow cytometric analysis of HeLa cells, untreated (blue) or IFN-α treated (green), using Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb compared to a nonspecific negative control antibody (red).
IF-IC
Confocal immunofluorescent analysis of HeLa cells, untreated (left) or IFNα-treated #9906 (right), using Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Chromatin IP
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HT-1080 cells treated with IFN-γ (50 ng/ml) for 30 minutes and either 5 μl of Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using human IRF-1 promoter primers, SimpleChIP® Human TAP1 Promoter Primers #5148, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
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
- Hebel, K. et al. (2011) J Immunol 187, 5627-35. Applications: Western Blotting
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Companion Products
- 9171 Phospho-Stat1 (Tyr701) Antibody
- 9172 Stat1 Antibody
- 9174 Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb (Alexa Fluor® 488 Conjugate)
- 8105 SignalSlide® Phospho-Stat1/3/5 IHC Controls
- 9170 PhosphoPlus® Stat1 (Tyr701) Antibody Kit
- 9173 Stat1 Control Cell Extracts
- 9175 Stat1 (42H3) Rabbit mAb
- 9176 Stat1 (9H2) Mouse mAb
- 6331 SignalSilence® Stat1 siRNA I
- 1038 Phospho-Stat1 (Tyr701) Blocking Peptide
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
- 8114 SignalStain® Boost IHC Detection Reagent (HRP, Rabbit)
- 8112 SignalStain® Antibody Diluent
- 8062 Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb (PE Conjugate)
- 8217 PhosphoPlus® Stat1 (Tyr701) Antibody Duet
Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.
For Research Use Only. Not For Use In Diagnostic Procedures.