Jak/Stat Pathway Inhibitors Antibody Sampler Kit

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Jak/Stat Pathway Inhibitors Antibody Sampler Kit

品牌： CST

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荧光素标记

Unconjugated

抗原名称

JakStat Pathway Inhibitors

简单描述

The Jak/Stat Pathway Inhibitors Antibody Sampler Kit provides an economical means to examine several inhibitors of Jak/Stat signaling, including PIAS1, PIAS3, PIAS4, SOCS1, SOCS2, and SOCS3. The kit contains enough primary antibody to perform two western blot experiments with each primary antibody.

研究领域

免疫学和肿瘤学,神经科学,

背景

Jak (Janus Kinase) and Stat (signal transducer and activator of transcription) proteins are utilized by receptors for a wide varity of ligands including cytokines, hormones, growth factors, and neurotransmitters (1). Jaks and Stats play important roles in oncogenesis, tumor progression, angiogenesis, cell motility, immune responses, and stem cell differentiation (2-5). Therefore, regulation of Jak/Stat signaling is crucial to prevent aberrant signaling which can lead to disease progression. Two methods for regulating Jak/Stat signaling involve SOCS and PIAS proteins (6,7).The SOCS (suppressor or cytokine signaling) family members are negative regulators of cytokine signal transduction that inhibit the Jak/Stat pathway and consist of 8 known members, including the originally identified protein CIS1 (cytokine-inducible SH2-containing protein) and SOCS1-SOCS7. Each SOCS family member contains a central SH2 domain and a conserved carboxy-terminal motif designated as the SOCS box. These proteins are important regulators of cytokine signaling, proliferation, differentiation, and immune responses (8-10). SOCS proteins are involved in regulating over 30 cytokines, including interleukins, growth hormone (GH), interferons, leptin, and leukemia inhibitory factor (7). SOCS1, also known as JAB (Janus Kinase binding protein) and SSI-1 (Stat-induced Stat inhibitor-1), shares the most homology with SOCS3 and both are highly induced by cytokines (7,11). Both SOCS1 and SOCS3 directly inhibit Jak activity; SOCS1 inhibits Jak through an interaction involving a phospohotyrosine located in the kinase activation loop; SOCS3 inhibits Jak via its SH2 domain (12,13). In addition to inhibiting Jak/Stat signaling, the SOCS box of SOCS1 and SOCS3 can trigger ubiquitin-mediated degradation of proteins within and outside the Jak/Stat pathway (14,15). SOCS2 is also incduced upon cytokine stimulation and the activity of SOCS2 has been predominately linked to GH and insulin-like growth fac 1.Darnell, J.E. et al. (1994) Science 264, 1415-21. 2.Bromberg, J.F. et al. (1999) Cell 98, 295-303. 3.Dentelli, P. et al. (1999) J Immunol 163, 2151-9. 4.Cattaneo, E. et al. (1999) Trends Neurosci 22, 365-9. 5.Su, L. et al. (1999) J Biol Chem 274, 31770-4. 6.Shuai, K. (2006) Cell Res 16, 196-202. 7.Croker, B.A. et al. (2008) Semin Cell Dev Biol 19, 414-22. 8.Alexander, W.S. et al. (1999) J Leukoc Biol 66, 588-92. 9.Chen, X.P. et al. (2000) Immunity 13, 287-90. 10.Hilton, D.J. et al. (1998) Proc Natl Acad Sci U S A 95, 114-9. 11.Starr, R. et al. (1997) Nature 387, 917-21. 12.Yasukawa, H. et al. (1999) EMBO J 18, 1309-20. 13.Sasaki, A. et al. (1999) Genes Cells 4, 339-51. 14.Kamizono, S. et al. (2001) J Biol Chem 276, 12530-8. 15.Rui, L. et al. (2002) J Biol Chem 277, 42394-8. 16.Dey, B.R. et al. (1998) J Biol Chem 273, 24095-101. 17.Liu, B. et al. (1998) Proc Natl Acad Sci U S A 95, 10626-31. 18.Chung, C.D. et al. (1997) Science 278, 1803-5. 19.Arora, T. et al. (2003) J Biol Chem 278, 21327-30. 20.Nishida, T. et al. (2007) Biochem J 405, 481-8. 21.Zhou, S. et al. (2008) J Biol Chem 283, 20137-48.

研究领域

免疫学和肿瘤学,神经科学,

翻译后修饰

unmodified

数据库链接

Entrez-Gene ID

8651,51588,8835,10401,9021,8554

UniProt ID

O15524,Q8N2W9,O14508,Q9Y6X2,O14543,O75925

参考图片

Confocal immunofluorescent analysis of A-204 (positive, left) and Hep G2 (low expression, right) cells using PIAS3 (D5F9) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).

Western blot analysis of extracts from K562, A204 and PC12 cells using SOCS2 Antibody.

Western blot analysis of extracts from various cell lines using PIAS1 (D33A7) XP® Rabbit mAb.

Confocal immunofluorescent analysis of AGS (left, high-expressing) and SW480 (right, low-expressing) using PIAS1 (D33A7) XP® Rabbit mAb (green), DyLight™ 650 Phalloidin #12956 (red), and DAPI #4083 (blue).

Western blot analysis of extracts from 293 and CCRF-CEM cells using PIAS4 (D2F12) Rabbit mAb.

Western blot analysis of extracts from Hep G2 and A-375 cell lines, untreated (-) or treated with Human Oncostatin M #5367 (hOSM; 20 ng/ml, 1 hr; +), using SOCS3 (D6E1T) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Western blot analysis of extracts from various cell lines using SOCS1 (E3Q4M) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). The absence of signal for SOCS1 in U266B1 cells has been found to be caused by aberrant hypermethylation of the SOCS1 gene and confirms the specificity of the antibody for SOCS1.

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

Western blot analysis of extracts from RD cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® PIAS3 siRNA I #9073 (+), or SignalSilence® PIAS3 siRNA II #9031 (+), using PIAS3 (D5F9) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). The PIAS3 (D5F9) XP® Rabbit mAb confirms silencing of PIAS3 expression, while the β-Actin (D6A8) Rabbit mAb is used as a loading control.

Confocal imunofluorescent analysis of HeLa cells using PIAS1 (D33A7) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing full-length human SOCS3 (+), using SOCS3 (D6E1T) Rabbit mAb.

Western blot analysis of extracts from various cell lines, untreated or treated with mINFα (10ng/ml, 24 hr), using SOCS1 (E3Q4M) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Western blot analysis of extracts from various cell lines using PIAS3 (D5F9) XP® Rabbit mAb.

Flow cytometric analysis of KARPAS 299 cells using PIAS1 (D33A7) XP® Rabbit mAb (solid line) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Western blot analysis of extracts from KARPAS-299 cells, untreated (-) or treated with the Jak3 inhibitor WHI-P154 (40 μM, overnight; +), using SOCS3 (D6E1T) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb (lower). KARPAS cell line source: Dr. Abraham Karpas at the University of Cambridge.

Western blot analysis of extracts from 293T cells, untransfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human SOCS1 protein (hSOCS1-Myc/DDK; +), using SOCS1 (E3Q4M) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing full-length human PIAS3 (hPIAS3; +), using PIAS3 (D5F9) XP® Rabbit mAb.

Western blot analysis of extracts from mouse bone marrow derived macrophage (mBMDM) cell extracts, untreated (-) or treated with Lipopolysaccharides #14011 (LPS; 50 ng/ml, 4 hr; +), using SOCS3 (D6E1T) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).

Immunoprecipitation of SOCS1 protein from L-540 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is SOCS1 (E3Q4M) Rabbit mAb. Western blot analysis was performed using SOCS1 (E3Q4M) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.

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货号：

8343T

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