Revision 11

#9139

Store at -20C

Stat3 (124H6) Mouse mAb
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.

Applications:
W, W-S, IP, IHC-Bond, IHC-P, IF-IC, FC-FP, ChIP, C&R

Reactivity:
H M R Mk

Sensitivity:
Endogenous

MW (kDa):
79, 86

Source/Isotype:
Mouse IgG2a kappa

UniProt ID:
#P40763

Entrez-Gene Id:
6774

Product Usage Information

For optimal ChIP 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.

The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.
Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
Immunoprecipitation 1:200
IHC Leica Bond 1:300 - 1:1200
Immunohistochemistry (Paraffin) 1:300 - 1:1200
Immunofluorescence (Immunocytochemistry) 1:800 - 1:3200
Flow Cytometry (Fixed/Permeabilized) 1:100 - 1:400
Chromatin IP 1:100
CUT&RUN 1:100

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.

For a carrier free (BSA and azide free) version of this product see product #83541.

Specificity/Sensitivity

Stat3 (124H6) Mouse mAb detects endogenous levels of total Stat3 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide centered around amino acid Gln692 of human Stat3.

Background

The Stat3 transcription factor is an important signaling molecule for many cytokines and growth factor receptors (1) and is required for murine fetal development (2). Research studies have shown that Stat3 is constitutively activated in a number of human tumors (3,4) and possesses oncogenic potential (5) and anti-apoptotic activities (3). Stat3 is activated by phosphorylation at Tyr705, which induces dimerization, nuclear translocation, and DNA binding (6,7). Transcriptional activation seems to be regulated by phosphorylation at Ser727 through the MAPK or mTOR pathways (8,9). Stat3 isoform expression appears to reflect biological function as the relative expression levels of Stat3α (86 kDa) and Stat3β (79 kDa) depend on cell type, ligand exposure, or cell maturation stage (10). It is notable that Stat3β lacks the serine phosphorylation site within the carboxy-terminal transcriptional activation domain (8).

Background References

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 nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

W: Western Blotting W-S: Simple Western™ IP: Immunoprecipitation IHC-Bond: IHC Leica Bond IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized) ChIP: Chromatin IP C&R: CUT&RUN

Cross-Reactivity Key

H: Human M: Mouse R: Rat Mk: Monkey

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.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意，否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件，除非书 CST 的合法授书代表以书面形式书独接受， 否书均被拒书，并且无效。

专品专有“专供研究使用”的专专或专似的专专声明， 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的， 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专，且专用于研专用途。将专品用于专断、专防或治专目的， 或专专售（专独或作专专成）或其他商专目的而专专专品，均需要 CST 的专独专可。客专：(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品，或使用专品制造任何商专专品，(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专，或使用专品开专任何与 CST 的专品或服专专争的专品或服专， (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专，(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品 ， (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

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 11

#9139

Stat3 (124H6) Mouse mAb

Cell Signaling Technology Logo
Western blot analysis of extracts from various human cell lines using Stat3 (124H6) Mouse mAb #9139 (upper) or β-Actin (8H10D10) Mouse mAb #3700 (lower).
Western Blotting Image 1: Stat3 (124H6) Mouse mAb
Western blot analysis of extracts from various rodent and monkey samples using Stat3 (124H6) Mouse mAb #9139 (upper) or β-Actin (8H10D10) Mouse mAb #3700 (lower).
Western Blotting Image 2: Stat3 (124H6) Mouse mAb
Simple Western™ analysis of lysates (1 mg/mL) from serum-starved HeLa cells treated with interferon-alpha (100 ng/mL, 5”) using Stat3 (124H6) Mouse mAb #9139. 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.
Western Blotting Image 1: Stat3 (124H6) Mouse mAb
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 11

#9139

Stat3 (124H6) Mouse mAb

Cell Signaling Technology Logo
Immunoprecipitation of Stat3 from HeLa cell extracts. Lane 1 is 10% input, lane 2 is precipitated with Mouse (G3A1) mAb IgG1 Isotype Control #5415, and lane 3 is Stat3 (124H6) Mouse mAb, #9139.
Immunoprecipitation Image 1: Stat3 (124H6) Mouse mAb
Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using Stat3 (124H6) Mouse mAb performed on the Leica BOND RX.
Immunohistochemistry Image 1: Stat3 (124H6) Mouse mAb
Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using Stat3 (124H6) Mouse mAb performed on the Leica BOND RX.
Immunohistochemistry Image 2: Stat3 (124H6) Mouse mAb
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 11

#9139

Stat3 (124H6) Mouse mAb

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma using Stat3 (124H6) Mouse mAb performed on the Leica BOND RX.
Immunohistochemistry Image 3: Stat3 (124H6) Mouse mAb
Immunohistochemical analysis of paraffin-embedded human papillary carcinoma of the breast using Stat3 (124H6) Mouse mAb performed on the Leica BOND RX.
Immunohistochemistry Image 4: Stat3 (124H6) Mouse mAb
Immunohistochemical analysis of paraffin-embedded human breast carcinoma (left), showing nuclear and cytoplasmic staining, and human lung carcinoma (right), showing cytoplasmic staining, using Stat3 (124H6) Mouse mAb.
Immunohistochemistry Image 1: Stat3 (124H6) Mouse mAb
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 11

#9139

Stat3 (124H6) Mouse mAb

Cell Signaling Technology Logo
Confocal immunofluorescent analysis of HeLa cells either serum-starved (left) or IFNalpha-treated (right) and labeled with Stat3 (124H6) Mouse mAb (green).
Immunofluorescence Image 1: Stat3 (124H6) Mouse mAb
Flow cytometric analysis of PC-3 cells (blue) and HeLa cells (green) using Stat3 (124H6) Mouse mAb (solid lines) or a concentration-matched Mouse (G3A1) mAb IgG1 Isotype Control #5415 (dashed lines). Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody.
Flow Cytometry Image 1: Stat3 (124H6) Mouse mAb
Chromatin immunoprecipitations were performed with cross-linked chromatin from Hep G2 cells starved overnight and treated with IL-6 (100 ng/ml) for 30 minutes, and either Stat3 (124H6) Mouse 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.
Chromatin Immunoprecipitation Image 1: Stat3 (124H6) Mouse mAb
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 11

#9139

Stat3 (124H6) Mouse mAb

Cell Signaling Technology Logo
CUT&RUN was performed with Hep G2 cells starved overnight and treated with IL-6 (100 ng/ml) for 30 minutes, and Stat3 (124H6) Mouse mAb, using CUT&RUN Assay Kit #86652. DNA Library was prepared using DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795. The figures show binding across BRD8 gene, a known target gene of Stat3 (see additional figure containing CUT&RUN-qPCR data).
CUT & RUN Image 1: Stat3 (124H6) Mouse mAb
CUT&RUN was performed with Hep G2 cells starved overnight and treated with IL-6 (100 ng/ml) for 30 minutes, and Stat3 (124H6) Mouse mAb, using CUT&RUN Assay Kit #86652. DNA Libraries were prepared using DNA Library Prep Kit for Illumina (ChIP-seq, CUT&RUN) #56795. The figures show binding across chromosome 5 (upper), including BRD8 gene (lower), a known target gene of Stat3 (see additional figure containing CUT&RUN-qPCR data).
CUT & RUN Image 2: Stat3 (124H6) Mouse mAb
CUT&RUN was performed with Hep G2 cells starved overnight and treated with IL-6 (100 ng/ml) for 30 minutes and either Stat3 (124H6) Mouse mAb or Rabbit (DA1E) mAb IgG XP® Isotype Control (CUT&RUN) #66362, using CUT&RUN Assay Kit #86652. The enriched DNA was quantified by real-time PCR using human BRD8 promoter primers and human PAIS3 upstream primers. 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.
CUT & RUN Image 3: Stat3 (124H6) Mouse mAb
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.