Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Western Blot, Intracellular Staining by Flow Cytometry

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 788335
View all STAT3 Primary Antibodies »

Product Specifications

Immunogen

Phosphopeptide containing the human STAT3 S727 site

Specificity

Detects human STAT3 when phosphorylated at S727.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Scientific Data Images for Human Phospho-STAT3 (S727) Antibody

Detection of Human Phospho-STAT3 (S727) antibody by Western Blot.

Detection of Human Phospho-STAT3 (S727) by Western Blot.

Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line untreated (-) or treated (+) with 100 ng/mL Recombinant Human IL-6 (Catalog # 206-IL) for 15 minutes. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Phospho-STAT3 (S727) Monoclonal Antibody (Catalog # MAB4934) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Phospho-STAT3 (S727) at approximately 95 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Phospho-STAT3 (S727) antibody in IFN alpha-treated Daudi Human Cell Line antibody by Flow Cytometry.

Detection of Phospho-STAT3 (S727) in IFN alpha-treated Daudi Human Cell Line by Flow Cytometry.

Daudi human Burkitt's lymphoma cell line was unstimulated (open histogram) or treated with 500 U/mL rhIFN-alpha for 20 minutes (filled histogram) and stained with Mouse anti-Human Phospho-STAT3 (S727) Molyclonal Antibody (Catalog # MAB4934) followed by APC-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with methanol. View our protocol for Staining Intracellular Molecules.

Detection of Human STAT3 by Western Blot

Detection of Human STAT3 by Western Blot

Regulation of hepcidin by IL-10 in primary macrophages, showing (a) the effect of IL-10 on Hepcidin mRNA synthesis with and without anti-IL-10 blocking antibody; (b) Western blot of STAT3 phosphorylation 20 minutes after exposure to increasing concentrations of IL-10; (c) STAT3-P inhibition of IL-10 (30 ng/mL) by Stattic.Bar plots show data from at least 3 independent experiments and the sho Western blot shows data from a representative experiment. M  =  media, iso ctrl  =  isotype control. * P<0.05, ***P<0.01 (Mann-Whitney test, compared with Media control). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/24520384), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human STAT3 by Western Blot

Detection of Human STAT3 by Western Blot

Regulation of hepcidin by IL-10 in primary macrophages, showing (a) the effect of IL-10 on Hepcidin mRNA synthesis with and without anti-IL-10 blocking antibody; (b) Western blot of STAT3 phosphorylation 20 minutes after exposure to increasing concentrations of IL-10; (c) STAT3-P inhibition of IL-10 (30 ng/mL) by Stattic.Bar plots show data from at least 3 independent experiments and the sho Western blot shows data from a representative experiment. M  =  media, iso ctrl  =  isotype control. * P<0.05, ***P<0.01 (Mann-Whitney test, compared with Media control). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/24520384), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Phospho-STAT3 (S727) by Western Blot

Detection of Phospho-STAT3 (S727) by Western Blot

Galectin-3 expression induces activation of PYK2, STAT1 and GSK3 alpha / beta signalling. Expression of 37 protein kinases in SW620 cells in response to 10 µg/ml galectin-3 or BSA for 0.5 h was assessed by Proteome Profiler Human Phospho-Kinase Array (A, Percentage changes of the kinases in cell response to galectin-3 in comparison to control are shown at the bottom panel). The presence of galectin-3 increases the phosphorylation of PYK2, GSK3 alpha / beta, and STAT1 and decreases phosphorylation of STAT3. SW620 cells treated with 10 µg/ml galectin-3 for different times were assessed by immunoblotting using antibodies against p-PYK2, p-STAT-1, p-GSK3 alpha / beta or p-STAT-3 (B). The blots were striped and reprobed with antibodies against PYK2, STAT-1, GSK3 alpha / beta or STAT-3. The band density was quantified and expressed as percentages of phospho-/non-phosphorylated proteins (C). In D and E, SW620 cells were treated with 10 µg/ml galectin-3 or BSA followed by introduction of GSK3 alpha / beta inhibitor SB 216763 (SB) or PKY2 inhibitor PF-431396 (PF) for 15 min and the levels of phosphorylated PYK2, STAT-1, GSK3 alpha / beta or STAT-3 were analysed by immunoblotting. The blots were striped and reprobed with antibodies against PYK2, STAT-1, GSK3 alpha / beta or STAT-3. The densities of the blots from three independent experiments were quantified and are expressed as the percentage of phosphorylated/non-phosphorylated levels of each protein. ***P < 0.001, **P < 0.01, *P < 0.05 (ANOVA). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37055381), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Phospho-STAT3 (S727) by Western Blot

Detection of Phospho-STAT3 (S727) by Western Blot

Galectin-3 expression induces activation of PYK2, STAT1 and GSK3 alpha / beta signalling. Expression of 37 protein kinases in SW620 cells in response to 10 µg/ml galectin-3 or BSA for 0.5 h was assessed by Proteome Profiler Human Phospho-Kinase Array (A, Percentage changes of the kinases in cell response to galectin-3 in comparison to control are shown at the bottom panel). The presence of galectin-3 increases the phosphorylation of PYK2, GSK3 alpha / beta, and STAT1 and decreases phosphorylation of STAT3. SW620 cells treated with 10 µg/ml galectin-3 for different times were assessed by immunoblotting using antibodies against p-PYK2, p-STAT-1, p-GSK3 alpha / beta or p-STAT-3 (B). The blots were striped and reprobed with antibodies against PYK2, STAT-1, GSK3 alpha / beta or STAT-3. The band density was quantified and expressed as percentages of phospho-/non-phosphorylated proteins (C). In D and E, SW620 cells were treated with 10 µg/ml galectin-3 or BSA followed by introduction of GSK3 alpha / beta inhibitor SB 216763 (SB) or PKY2 inhibitor PF-431396 (PF) for 15 min and the levels of phosphorylated PYK2, STAT-1, GSK3 alpha / beta or STAT-3 were analysed by immunoblotting. The blots were striped and reprobed with antibodies against PYK2, STAT-1, GSK3 alpha / beta or STAT-3. The densities of the blots from three independent experiments were quantified and are expressed as the percentage of phosphorylated/non-phosphorylated levels of each protein. ***P < 0.001, **P < 0.01, *P < 0.05 (ANOVA). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37055381), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Phospho-STAT3 (S727) by Western Blot

Detection of Phospho-STAT3 (S727) by Western Blot

Galectin-3 expression induces activation of PYK2, STAT1 and GSK3 alpha / beta signalling. Expression of 37 protein kinases in SW620 cells in response to 10 µg/ml galectin-3 or BSA for 0.5 h was assessed by Proteome Profiler Human Phospho-Kinase Array (A, Percentage changes of the kinases in cell response to galectin-3 in comparison to control are shown at the bottom panel). The presence of galectin-3 increases the phosphorylation of PYK2, GSK3 alpha / beta, and STAT1 and decreases phosphorylation of STAT3. SW620 cells treated with 10 µg/ml galectin-3 for different times were assessed by immunoblotting using antibodies against p-PYK2, p-STAT-1, p-GSK3 alpha / beta or p-STAT-3 (B). The blots were striped and reprobed with antibodies against PYK2, STAT-1, GSK3 alpha / beta or STAT-3. The band density was quantified and expressed as percentages of phospho-/non-phosphorylated proteins (C). In D and E, SW620 cells were treated with 10 µg/ml galectin-3 or BSA followed by introduction of GSK3 alpha / beta inhibitor SB 216763 (SB) or PKY2 inhibitor PF-431396 (PF) for 15 min and the levels of phosphorylated PYK2, STAT-1, GSK3 alpha / beta or STAT-3 were analysed by immunoblotting. The blots were striped and reprobed with antibodies against PYK2, STAT-1, GSK3 alpha / beta or STAT-3. The densities of the blots from three independent experiments were quantified and are expressed as the percentage of phosphorylated/non-phosphorylated levels of each protein. ***P < 0.001, **P < 0.01, *P < 0.05 (ANOVA). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37055381), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Phospho-STAT3 (S727) by Western Blot

Detection of Phospho-STAT3 (S727) by Western Blot

Galectin-3 expression induces activation of PYK2, STAT1 and GSK3 alpha / beta signalling. Expression of 37 protein kinases in SW620 cells in response to 10 µg/ml galectin-3 or BSA for 0.5 h was assessed by Proteome Profiler Human Phospho-Kinase Array (A, Percentage changes of the kinases in cell response to galectin-3 in comparison to control are shown at the bottom panel). The presence of galectin-3 increases the phosphorylation of PYK2, GSK3 alpha / beta, and STAT1 and decreases phosphorylation of STAT3. SW620 cells treated with 10 µg/ml galectin-3 for different times were assessed by immunoblotting using antibodies against p-PYK2, p-STAT-1, p-GSK3 alpha / beta or p-STAT-3 (B). The blots were striped and reprobed with antibodies against PYK2, STAT-1, GSK3 alpha / beta or STAT-3. The band density was quantified and expressed as percentages of phospho-/non-phosphorylated proteins (C). In D and E, SW620 cells were treated with 10 µg/ml galectin-3 or BSA followed by introduction of GSK3 alpha / beta inhibitor SB 216763 (SB) or PKY2 inhibitor PF-431396 (PF) for 15 min and the levels of phosphorylated PYK2, STAT-1, GSK3 alpha / beta or STAT-3 were analysed by immunoblotting. The blots were striped and reprobed with antibodies against PYK2, STAT-1, GSK3 alpha / beta or STAT-3. The densities of the blots from three independent experiments were quantified and are expressed as the percentage of phosphorylated/non-phosphorylated levels of each protein. ***P < 0.001, **P < 0.01, *P < 0.05 (ANOVA). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37055381), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human Phospho-STAT3 (S727) Antibody

Application
Recommended Usage

Intracellular Staining by Flow Cytometry

0.25 µg/106 cells
Sample: IFN alpha-treated Daudi Human Cell Line fixed with paraformaldehyde and permeabilized with methanol

Western Blot

1 µg/mL
Sample: HepG2 human hepatocellular carcinoma cell line treated with Recombinant Human IL‑6 (Catalog # 206-IL)

Reviewed Applications

Read 1 review rated 5 using MAB4934 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: STAT3

Signal Transducer and Activator of Transcription (STAT) proteins are transcription factors activated in response to cytokine, growth factor, or hormone receptor signaling. Janus kinases (JAKs) phosphorylate STAT proteins and induce dimerization. Homo- or heterodimers translocate to the nucleus where they bind to DNA and activate transcription.

Long Name

Signal Transducer and Activator of Transcription 3

Alternate Names

Acute-phase response factor, APRFMGC16063, DNA-binding protein APRF, FLJ20882, HIES, signal transducer and activator of transcription 3, signal transducer and activator of transcription 3 (acute-phase responsefactor)

Entrez Gene IDs

6774 (Human); 20848 (Mouse); 25125 (Rat)

Gene Symbol

STAT3

Additional STAT3 Products

Product Documents for Human Phospho-STAT3 (S727) Antibody

Certificate of Analysis

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Product Specific Notices for Human Phospho-STAT3 (S727) Antibody

For research use only

Citations for Human Phospho-STAT3 (S727) Antibody

Customer Reviews for Human Phospho-STAT3 (S727) Antibody (1)

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  • Human Phospho-STAT3 (S727) Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Human breast cancer cell lines
    Species: Human
    Verified Customer | Posted 10/11/2018
    Human Phospho-STAT3 (S727) Antibody MAB4934

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