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8113
SignalStain® Phospho-Stat IHC Sampler Kit
Primary Antibodies
IHC Kit

SignalStain® Phospho-Stat IHC Sampler Kit #8113

Citations (0)
SignalStain® Phospho-Stat IHC Sampler Kit: Image 1
Immunohistochemical analysis of paraffin-embedded human breast carcinoma comparing SignalStain® Antibody Diluent (left) to TBST/normal goat serum (right) using Phospho-Akt (Ser473) (D9E) Rabbit mAb #4060.
SignalStain® Phospho-Stat IHC Sampler Kit: Image 2
Chromatin immunoprecipitations were performed with cross-linked chromatin from Hep G2 cells starved overnight and treated with IL-6 (100 ng/ml) for 30 minutes and Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across IRF1, a known target gene of Phospho-Sata3 (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.
SignalStain® Phospho-Stat IHC Sampler Kit: Image 3
Western blot analysis of extracts from IFN-alpha treated Jurkat cells and HeLa cells (left), as well as EGF treated A431 cells (right), using Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb. Note that the basal phospho-Stat3 in A431 is detected by the antibody.
SignalStain® Phospho-Stat IHC Sampler Kit: Image 4
Chromatin immunoprecipitations were performed with cross-linked chromatin from HaCaT cells Hep G2 cells starved overnight and treated with IL-6 (100 ng/ml) for 30 minutes and either Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Sonication Chromatin IP Kit #56383. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human c-Fos Promoter Primers #4663, human IRF1 promoter primers, 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.
SignalStain® Phospho-Stat IHC Sampler Kit: Image 5
Immunohistochemical analysis of paraffin-embedded human neuroendocrine lung carcinoma using Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb performed on the Leica® BOND™ Rx.
SignalStain® Phospho-Stat IHC Sampler Kit: Image 6
Chromatin immunoprecipitations were performed with cross-linked chromatin from Hep G2 cells starved overnight and treated with IL-6 (100 ng/ml) for 30 minutes, and either Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb or 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 c-Fos Promoter Primers #4663, 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.
SignalStain® Phospho-Stat IHC Sampler Kit: Image 7
Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb performed on the Leica® BOND™ Rx.
SignalStain® Phospho-Stat IHC Sampler Kit: Image 8
Immunohistochemical analysis of paraffin-embedded Apc (min/+) mouse intestine, using Phosho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb.
SignalStain® Phospho-Stat IHC Sampler Kit: Image 9
Immunohistochemical analysis of paraffin embedded human breast carcinoma, specifically endothelial cells, untreated (left) or lambda phosphatase treated (right), using Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb.
SignalStain® Phospho-Stat IHC Sampler Kit: Image 10
Immunnohistochemical analysis of paraffin-embedded human lung carcinoma, showing nuclear localization, using Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb.
SignalStain® Phospho-Stat IHC Sampler Kit: Image 11
Immunohistochemical analysis using Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb on SignalSlide® HeLa -/+ IFNa IHC Controls #55861 (paraffin-embedded HeLa cell pellets, untreated (left) or treated with Human Interferon-α1 (hIFN-α1) #8927 (right)).
SignalStain® Phospho-Stat IHC Sampler Kit: Image 12
Confocal immunofluorescent analysis of HeLa cells, IFN-alpha treated (left) or untreated (right), labeled with Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb (green).
SignalStain® Phospho-Stat IHC Sampler Kit: Image 13
Flow cytometric analysis of U266 cells, untreated (blue) or treated with IFNalpha (50 ng/ml, 15 min; green) using Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb (solid lines) or concentration matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
SignalStain® Phospho-Stat IHC Sampler Kit: Image 14
Chromatin immunoprecipitations were performed with cross-linked chromatin from HT-1080 cells treated with IFN-γ (50 ng/ml) for 30 minutes and Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across IRF-1, a known target gene of Phospho-Stat1 (see additional figure containing ChIP-qPCR data).
SignalStain® Phospho-Stat IHC Sampler Kit: Image 15
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).
SignalStain® Phospho-Stat IHC Sampler Kit: Image 16
Chromatin immunoprecipitations were performed with cross-linked chromatin from HT-1080 cells treated with IFN-γ (50 ng/ml) for 30 minutes and Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across chromosome 5 (upper), including IRF-1 (lower), a known target gene of Phospho-Stat1 (see additional figure containing ChIP-qPCR data).
SignalStain® Phospho-Stat IHC Sampler Kit: Image 17
Immunohistochemical analysis of paraffin-embedded Non-Hodgkin's lymphoma control (left) or λ phosphatase treated (right), using Phospho-Stat1 (tyr701) (58D6) Rabbit mAb.
SignalStain® Phospho-Stat IHC Sampler Kit: Image 18
Chromatin immunoprecipitations were performed with cross-linked chromatin from HT-1080 cells treated with IFN-γ (50 ng/ml) for 30 minutes and either Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. 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.
SignalStain® Phospho-Stat IHC Sampler Kit: Image 19
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).
SignalStain® Phospho-Stat IHC Sampler Kit: Image 20
Immunohistochemical analysis of paraffin-embedded stomach (chronic gastritis), using Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb.
SignalStain® Phospho-Stat IHC Sampler Kit: Image 21
Immunohistochemical analysis using Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb on SignalSlide® HeLa -/+ IFNa IHC Controls #55861 (paraffin-embedded HeLa cell pellets, untreated (left) or treated with Human Interferon-α1 (hIFN-α1) #8927 (right)).
SignalStain® Phospho-Stat IHC Sampler Kit: Image 22
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).
SignalStain® Phospho-Stat IHC Sampler Kit: Image 23
Flow cytometric analysis of Jurkat cells, untreated (blue) or treated with IFN-α (100ng/ml, 5 mins; green) using Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412.
SignalStain® Phospho-Stat IHC Sampler Kit: Image 24
Western blot analysis of extracts from K562 cells, untreated or treated with Gleevec, using Phospho-Stat5 (Tyr694) (C11C5) Rabbit mAb.
SignalStain® Phospho-Stat IHC Sampler Kit: Image 25
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-Stat5 (Tyr694) (C11C5) Rabbit mAb.
SignalStain® Phospho-Stat IHC Sampler Kit: Image 26
Flow cytometric analysis of K562 cells, untreated (green) or STI571-treated (blue) using Phospho-Stat5 (Tyr694) (C11C5) Rabbit mAb.
Inquiry Info.# 8113

Important Ordering Details

Custom Ordering Details: The control slides that accompany this kit are cut freshly upon ordering. Please allow up to three business days for your product to be processed.

Product Description

The SignalStain® Phospho-Stat IHC Sampler Kit from Cell Signaling Technology allows the researcher to examine paraffin-embedded tissues or cells with antibodies that will detect phosphorylated Stat proteins. Each antibody is validated for use in immunohistochemical assays using multiple approaches. Also included in the kit are control slides that can be used to verify the performance of each antibody and a primary antibody diluent. Please see table above for the recommended antibody diluent for each antibody provided in the kit.

Specificity / Sensitivity

Each antibody in the SignalStain® Phospho-Stat IHC Sampler Kit detects endogenous levels of its target protein. Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb detects endogenous levels of Stat1 only when phosphorylated at Tyr701; p91 Stat1 phosphorylated at Tyr701 and the p84 splice variant will also be detected by this antibody. It does not cross-react with the corresponding phospho-tyrosines of other Stat proteins. Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb detects endogenous levels of Stat3 only when phosphorylated at Tyr705. This antibody does not cross-react with phospho-EGFR or the corresponding phospho-tyrosines of other Stat proteins. Phospho-Stat5 (Tyr694) (C11C5) Rabbit mAb detects endogenous Stat5a only when phosphorylated at Tyr694 and Stat5b when phosphorylated at Tyr699.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr701 of human Stat1, Tyr705 of mouse Stat3, or Tyr694 of Stat5a.

Background

Stat proteins serve as transcription factors in growth and survival pathways stimulated by growth factor and cytokine activation of receptor proteins. Receptor activation promotes tyrosine phosphorylation of Stat proteins, resulting in Stat dimerization and translocation to the nucleus where they regulate expression of numerous proteins that control cell growth, survival, differentiation and pathogen resistance (1). Stat1 is essential in IFN-α and IFN-γ stimulated pathways and is abnormally activated in many tumors (2,3). Both Stat1α (91 kDa) and Stat1β (84 kDa) isoforms are activated by IFN-α but only Stat1α responds to IFN-γ. Phosphorylation of Stat1 at Tyr701 induces Stat1 dimerization, nuclear translocation and DNA binding (4). Transcription factor Stat3 possesses oncogenic potential and anti-apoptotic activities; a number of human tumors display constitutively activated Stat3 (5,6). Activation of Stat3 follows phosphorylation at Tyr705, resulting in dimerization, nuclear translocation and DNA binding (7). Expression of Stat3α (86 kDa) and Stat3β (79 kDa) isoforms correlates with cell type, ligand and cell maturation stage (8). Phosphorylation of Stat5a at Tyr694 is essential for transcription factor activation (9). Stat5 is activated by interleukins and other cytokines (i.e. CSF1); presence of phosphorylated Stat5 is in some IL-3-treated cells suggests a role in angiogenesis and cell motility (10). Altered Stat5 expression and activity is associated with abnormal cell proliferation and apoptosis in some forms of leukemia (11).
  1. Ihle, J.N. (2001) Curr Opin Cell Biol 13, 211-7.
  2. Bromberg, J. (2002) J Clin Invest 109, 1139-42.
  3. Frank, D.A. (1999) Mol Med 5, 432-56.
  4. Ihle, J.N. et al. (1994) Trends Biochem Sci 19, 222-7.
  5. Garcia, R. and Jove, R. (1998) J Biomed Sci 5, 79-85.
  6. Catlett-Falcone, R. et al. (1999) Immunity 10, 105-15.
  7. Darnell, J.E. (1997) Science 277, 1630-5.
  8. Biethahn, S. et al. (1999) Exp Hematol 27, 885-94.
  9. Gouilleux, F. et al. (1994) EMBO J 13, 4361-9.
  10. Buitenhuis, M. et al. (2004) Int J Biochem Cell Biol 36, 2120-4.
  11. Baśkiewicz-Masiuk, M. et al. (2003) Cell Prolif 36, 265-78.

Pathways & Proteins

Explore pathways + proteins related to this product.

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For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
SignalStain is a trademark of Cell Signaling Technology, Inc.