Revision 4

#8343

Store at -20C

Jak/Stat Pathway Inhibitors Antibody Sampler Kit

1 Kit

(6 x 20 microliters)

CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

Web:

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Product Includes Product # Quantity Mol. Wt Isotype/Source
SOCS1 (E3Q4M) Rabbit Monoclonal Antibody5531320 µl23 kDaRabbit IgG
SOCS2 Antibody277920 µl22 kDaRabbit
SOCS3 (D6E1T) Rabbit Monoclonal Antibody5211320 µl28 kDaRabbit IgG
PIAS1 (D33A7) Rabbit Monoclonal Antibody355020 µl76 kDaRabbit IgG
PIAS3 (D5F9) Rabbit Monoclonal Antibody904220 µl65-75 kDaRabbit IgG
PIAS4 (D2F12) Rabbit Monoclonal Antibody439220 µl75 kDaRabbit IgG
Anti-rabbit IgG, HRP-linked Antibody7074100 µlGoat

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

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.

Storage

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.

Background

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

Background References

  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.

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 4

#8343

Jak/Stat Pathway Inhibitors Antibody Sampler Kit

Cell Signaling Technology Logo
Western blot analysis of extracts from K562, A204 and PC12 cells using SOCS2 Antibody.
undefined Image 1: Jak/Stat Pathway Inhibitors Antibody Sampler Kit
Western blot analysis of extracts from various cell lines using PIAS1 (D33A7) XP® Rabbit mAb.
undefined Image 2: Jak/Stat Pathway Inhibitors Antibody Sampler Kit
Western blot analysis of extracts from 293 and CCRF-CEM cells using PIAS4 (D2F12) Rabbit mAb.
undefined Image 3: Jak/Stat Pathway Inhibitors Antibody Sampler Kit
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 4

#8343

Jak/Stat Pathway Inhibitors Antibody Sampler Kit

Cell Signaling Technology Logo
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).
undefined Image 4: Jak/Stat Pathway Inhibitors Antibody Sampler Kit
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.
undefined Image 5: Jak/Stat Pathway Inhibitors Antibody Sampler Kit
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.
undefined Image 6: Jak/Stat Pathway Inhibitors Antibody Sampler Kit
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 4

#8343

Jak/Stat Pathway Inhibitors Antibody Sampler Kit

Cell Signaling Technology Logo
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.
undefined Image 7: Jak/Stat Pathway Inhibitors Antibody Sampler Kit
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.
undefined Image 8: Jak/Stat Pathway Inhibitors Antibody Sampler Kit
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).
undefined Image 9: Jak/Stat Pathway Inhibitors Antibody Sampler Kit
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 4

#8343

Jak/Stat Pathway Inhibitors Antibody Sampler Kit

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using PIAS3 (D5F9) XP® Rabbit mAb.
undefined Image 10: Jak/Stat Pathway Inhibitors Antibody Sampler Kit
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.
undefined Image 11: Jak/Stat Pathway Inhibitors Antibody Sampler Kit
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.
undefined Image 12: Jak/Stat Pathway Inhibitors Antibody Sampler Kit
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 4

#8343

Jak/Stat Pathway Inhibitors Antibody Sampler Kit

Cell Signaling Technology Logo
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).
undefined Image 13: Jak/Stat Pathway Inhibitors Antibody Sampler Kit
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.
undefined Image 14: Jak/Stat Pathway Inhibitors Antibody Sampler Kit
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).
undefined Image 15: Jak/Stat Pathway Inhibitors Antibody Sampler Kit
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 4

#8343

Jak/Stat Pathway Inhibitors Antibody Sampler Kit

Cell Signaling Technology Logo
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.
undefined Image 16: Jak/Stat Pathway Inhibitors Antibody Sampler Kit
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).
undefined Image 17: Jak/Stat Pathway Inhibitors Antibody Sampler Kit
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).
undefined Image 18: Jak/Stat Pathway Inhibitors Antibody Sampler Kit
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.