Revision 1

#9939

Store at -20C

Stat Antibody Sampler Kit

1 Kit

(4 x 20 microliters)

CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

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
Stat1 Antibody917220 µl84, 91 kDaRabbit
Stat3 (79D7) Rabbit mAb490420 µl79, 86 kDaRabbit IgG
Stat5 (D2O6Y) Rabbit mAb9420520 µl90 kDaRabbit IgG
Stat6 (D3H4) Rabbit mAb539720 µl110 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 Stat Antibody Sampler Kit provides an economical means to examine multiple Stat proteins: Stat1, Stat3, Stat5 and Stat6. The kit contains enough primary and secondary antibodies to perform two Western blot experiments.

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

Janus kinases (Jaks) and signal transducers and activators of transcription (Stats) are utilized by receptors for a wide variety of ligands including cytokines, hormones, growth factors, and neurotransmitters. Jaks, activated via autophosphorylation following ligand-induced receptor aggregation, phosphorylate tyrosine residues on associated receptors, Stat molecules, and other downstream signaling proteins (1,2). The phosphorylation of Stat proteins at conserved tyrosine residues activates SH2-mediated dimerization followed rapidly by nuclear translocation. Stat dimers bind to interferon response element (IRE) and gamma interferon-activated sequence (GAS) DNA elements, resulting in the transcriptional regulation of downstream genes (1,2). The remarkable range and specificity of responses regulated by the Stats is determined in part by the tissue-specific expression of different cytokine receptors, Jaks and Stats (2,3), and by the combinatorial coupling of various Stat members to different receptors. Serine phosphorylation in the carboxy-terminal transcriptional activation domain has been shown to regulate the function of Stat1, Stat2, Stat3, Stat4, and Stat5 (1). Phosphorylation of Stat3 at Ser727 via MAPK or mTOR pathways is required for optimal transcriptional activation in response to growth factors and cytokines including IFN-gamma and ciliary neurotrophic factor (CNTF) (4,5). Jak/Stat pathways also play important roles in oncogenesis, tumor progression, angiogenesis, cell motility, immune responses, and stem cell differentiation (6-11).

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 1

Cell Signaling Technology Logo
Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® Stat3 siRNA I (+) or SignalSilence® Stat3 siRNA II #6582 (+), using Stat3 (79D7) Rabbit mAb #4904 and α-Tubulin (11H10) Rabbit mAb #2125. The Stat3 (79D7) Rabbit mAb confirms silencing of Stat3 expression, while the α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of Stat3 siRNA.
undefined Image 1: Stat Antibody Sampler Kit
Western blot analysis of extracts from various cell lines using Stat6 (D3H4) Rabbit mAb.
undefined Image 2: Stat 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 3: Stat 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 1

Cell Signaling Technology Logo
Western blot analysis of extracts from A549 cells (lane 1) or STAT1 knock-out cells (lane 2) using Stat1 Antibody #9172 (upper), and β-actin (D6A8) Rabbit mAb #8457 (lower). The absence of signal in the STAT1 knock-out A549 cells confirms specificity of the antibody for STAT1.
undefined Image 4: Stat Antibody Sampler Kit
Western blot analysis of extracts from various cell lines using Stat5 (D2O6Y) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
undefined Image 5: Stat Antibody Sampler Kit
Western blot analysis of extracts from various cell lines using Stat3 (79D7) Rabbit mAb.
undefined Image 6: Stat 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 1

Cell Signaling Technology Logo
Chromatin immunoprecipitations were performed with cross-linked chromatin from Ramos cells starved overnight then treated with IL-4 (100 ng/ml, 30 min) and either Stat6 (D3H4) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human DMD Intron 2 Primers #7710, human MS4A1 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.
undefined Image 7: Stat Antibody Sampler Kit
Western blot analysis of extracts from HeLa cells 48 hours following mock transfection, transfection with non-targeted (control) siRNA or transfection with Stat1 siRNA. Stat1 was detected using Stat1 Antibody #9172, and p42 was detected using p42 MAPK Antibody #9108. The Stat1 Antibody confirms silencing of Stat1 expression, and the p42 MAPK Antibody is used to control for protein loading and siRNA specificity.
undefined Image 8: Stat Antibody Sampler Kit
Western blot analysis of PC-3 cells, mock transfected (-) or transfected with constructs expressing full-length human Stat5a (+) or Stat5b (+) using Stat5 (D2O6Y) Rabbit mAb.
undefined Image 9: Stat 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 1

Cell Signaling Technology Logo
Western blot analysis of extracts from SK-MEL-28 cells, untreated or IFN-alpha-treated (100 ng/ml), using Phospho-Stat1 (Tyr701) Antibody #9171 (upper) or Stat1 Antibody (lower).
undefined Image 10: Stat Antibody Sampler Kit
Immunoprecipitation of Stat5 from K-562 cell extracts. Lane 1 is 10% input, lane 2 is precipitated with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Stat5 (D2O6Y) Rabbit mAb. Western blot was performed using Stat5 (D2O6Y) Rabbit mAb.
undefined Image 11: Stat Antibody Sampler Kit
Chromatin immunoprecipitations were performed with cross-linked chromatin from Hep G2 cells treated with IL-6 (100 ng/ml) for 30 minutes, and either Stat3 (79D7) Rabbit mAb or 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.
undefined Image 12: Stat 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 1

Cell Signaling Technology Logo
Chromatin immunoprecipitations were performed with cross-linked chromatin from HT-1080 cells treated with IFN-γ (50 ng/ml) for 30 minutes and either Stat1 Antibody or 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.
undefined Image 13: Stat Antibody Sampler Kit
Chromatin immunoprecipitations were performed with cross-linked chromatin from BaF3 cells starved of IL-3 for 6 hours followed by induction with IL-3 for 45 minutes, and either Stat5 (D2O6Y) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Mouse CIS Intron 1 Primers #5131, mouse SOCS-3 promoter primers, and SimpleChIP® Mouse RPL30 Intron 2 Primers #7015. 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.
undefined Image 14: Stat Antibody Sampler Kit
Simple WesternTM analysis of lysates (0.1 mg/mL) from 3T3 cells using Stat3 (79D7) Rabbit mAb #4904. The virtual lane view (left) shows the target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody.  The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (green line) dilutions of primary antibody.  This experiment was performed under reducing conditions on the JessTM Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
undefined Image 15: Stat 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 1

Cell Signaling Technology Logo

Simple Western™ analysis of lysates (0.1 mg/mL) from Jurkat cells using Stat6 (D3H4) Rabbit mAb #5397. The virtual lane view (left) shows the 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.

undefined Image 16: Stat Antibody Sampler Kit
Simple Western™ analysis of lysates (0.1 mg/mL) from HDLM-2 cells using Stat5 (D2O6Y) Rabbit mAb #94205. The virtual lane view (left) shows the target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (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.
undefined Image 17: Stat 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.