Revision 3

#2653Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, ChIP, ChIP-seq

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

81

Source/Isotype:

Rabbit 

UniProt ID:

#Q14765

Entrez-Gene Id:

6775

Product Information

Product Usage Information

For optimal ChIP and ChIP-seq results, use 10 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.

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Chromatin IP 1:50
Chromatin IP-seq 1:50

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.

Specificity / Sensitivity

Stat4 (C46B10) Rabbit mAb detects endogenous levels of total Stat4 protein.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Lys151 of Stat4.

Background

The Jak-Stat signaling pathway is utilized by a large number of cytokines, growth factors, and hormones (1). Receptor-mediated tyrosine phosphorylation of Jak family members triggers phosphorylation of Stat proteins, resulting in their nuclear translocation, binding to specific DNA elements, and subsequent activation of transcription. 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, as well as by the combinatorial coupling of various Stat members to different receptors (2). Stat4 is predominantly expressed in the spleen, thymus, and testis and has been most extensively investigated as the mediator of IL-12 responses (3-8). Activation of Stat4 is associated with phosphorylation at Tyr693 (9).

  1. Darnell, J.E. et al. (1994) Science 264, 1415-1421.
  2. Leonard, W.J. and O'Shea, J.J. (1998) Annu. Rev. Immunol. 16, 293-322.
  3. Zhong, Z. et al. (1994) Proc. Natl. Acad. Sci. USA 91, 4806-4810.
  4. Yamamoto, K. et al. (1994) Mol. Cell Biol. 14, 4342-4349.
  5. Jacobson, N.G. et al. (1995) J. Exp. Med. 181, 1755-1762.
  6. Bacon, C.M. et al. (1995) Proc. Natl. Acad. Sci. USA 92, 7307-7311.
  7. Thierfelder, W.E. et al. (1996) Nature 382, 171-174.
  8. Kaplan, M.H. et al. (1996) Nature 382, 174-177.
  9. Visconti, R. et al. (2000) Blood 96, 1844-52.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
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.

Revision 3
#2653

Stat4 (C46B10) Rabbit mAb

Western Blotting Image 1: Stat4 (C46B10) Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines and from rat thymus using Stat4 (C46B10) Rabbit mAb.
Western Blotting Image 2: Stat4 (C46B10) Rabbit mAb Expand Image
Western blot analysis of extracts from ACHN, SR and Caki cell lines and from rat thymus using Stat4 (C46B10) Rabbit mAb.
Immunoprecipitation Image 1: Stat4 (C46B10) Rabbit mAb Expand Image
Immunoprecipitation of Stat4 protein from NK-92 cell extracts. Lane 1 is 10% input, lane 2 is beads only, lane 3 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 4 is Stat4 (C46B10) Rabbit mAb. Western blot analysis was performed using Stat4 (C46B10) Rabbit mAb.
Chromatin Immunoprecipitation Image 1: Stat4 (C46B10) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from NK-92 cells starved of IL-2 overnight then treated with IL-12 (10 ng/ml) for 4 hr and Stat4 (C46B10) 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 IRF1, a known target gene of Stat4 (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product datasheet.
Chromatin Immunoprecipitation Image 2: Stat4 (C46B10) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from NK-92 cells starved of IL-2 overnight then treated with IL-12 (10 ng/ml) for 4 hr and Stat4 (C46B10) 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 IRF1 (lower), a known target gene of Stat4 (see additional figure containing ChIP-qPCR data).
Chromatin Immunoprecipitation Image 3: Stat4 (C46B10) Rabbit mAb Expand Image
Chromatin immunoprecipitations were performed with cross-linked chromatin from NK-92 cells starved of IL-2 overnight then treated with IL-12 (10 ng/ml) for 4 hr and either Stat4 (C46B10) Rabbit mAb #2653 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, human PRF1 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.