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Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb
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Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb

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分子量:
84, 91
来源宿主:
Rabbit
产品介绍
产品介绍
产品信息
荧光素标记
抗原名称
Stat1
来源纯化

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr701 of human Stat1.

宿主
Rabbit
商品描述

Product Usage Information

For optimal ChIP and ChIP-seq results, use 5 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

ApplicationDilution
Western Blotting1:1000
Simple Western™1:10 - 1:50
Immunoprecipitation1:100
Immunohistochemistry (Paraffin)1:400 - 1:1600
Immunofluorescence (Immunocytochemistry)1:200 - 1:800
Flow Cytometry (Fixed/Permeabilized)1:100 - 1:400
Chromatin IP1:100
Chromatin IP-seq1:100
同种型
Rabbit IgG
分子量
84, 91
研究领域
免疫学和肿瘤学,神经科学,
应用
反应种属
Human,Mouse,
目标/特异性

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.

Species Reactivity:

Human, Mouse

敏感性
Endogenous
背景
背景
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. 1.Heim, M.H. (1999) J Recept Signal Transduct Res 19, 75-120. 2.Durbin, J.E. et al. (1996) Cell 84, 443-50. 3.Meraz, M.A. et al. (1996) Cell 84, 431-42. 4.Ihle, J.N. et al. (1994) Trends Biochem Sci 19, 222-7. 5.Frank, D.A. (1999) Mol Med 5, 432-56. 6.Wen, Z. et al. (1995) Cell 82, 241-50.
研究领域
免疫学和肿瘤学,神经科学,
翻译后修饰
phosphate
制备和贮存
保存方式

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier-free (BSA and azide free) version of this product see product #88845.

数据库链接
Entrez-Gene ID
6772
UniProt ID
P42224

参考图片

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.

Western blot analysis of extracts from HeLa cells, untreated or treated with IFNa (#36000, 100 ng/mL, 5 min) or IFNg (#80385, 100 ng/mL, 30 min); using Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb #9167 (upper), Stat1 (D1K9Y) Rabbit mAb #14994 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Western blot analysis of extracts from various cell lines, untreated or treated with EGF (100 ng/mL, 30 min), IFNa (100 ng/mL, 5 min), or PDGF (100 ng/mL, 5 min); using Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb #9167 (upper), and β-Actin (D6A8) Rabbit mAb #8457 (lower).

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).

Simple Western™ analysis of lysates (0.1 mg/mL) from serum-starved HeLa cells treated with IFN-alpha (100 ng/mL, 5 min) using Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb #9167. The virtual lane view (left) shows a single target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.

Immunoprecipitation of Phospho-Stat1 (Tyr701) from HeLa cell extracts, treated with treated with IFNa (#36000, 100 ng/mL, 5 min). Lane 1 is 10% input, lane 2 is precipitated with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb, #9167.

Immunohistochemical analysis of paraffin-embedded Non-Hodgkin lymphoma control (left) or λ phosphatase treated (right), using Phospho-Stat1 (tyr701) (58D6) Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded Non-Hodgkin lymphoma using Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb #9167 in the presence of control peptide (left) or Phospho-Stat1 (Tyr701) Blocking Peptide (right).

Immunohistochemical analysis of paraffin-embedded stomach (chronic gastritis), using Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb.

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)).

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).

Flow cytometric analysis of U266B1 cells, untreated (blue) or treated with hIFN-β (100 ng/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.

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.

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 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across IRF-1, a known target gene of Phospho-Stat1 (see additional figure containing ChIP-qPCR data).

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 DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #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).

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货号:
9167T
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