STAT3 Antibody (9D8)

STAT3 Monoclonal Antibody for Western blot, IF, IHC (P), IP, ChIP

>> See 17 other antibodies for STAT3 (APRF; HIES; FLJ20882; MGC16063; STAT3, Signal transducer and activator of transcription 3)
Synonyms:
APRF; HIES; FLJ20882; MGC16063; STAT3, Signal transducer and activator of transcription 3
Entrez Gene ID:
Details
Host / Isotype: Mouse / IgG
Class: Monoclonal
Type: Antibody
Clone: 9D8
Tested Species Reactivity: Human (Hu), Mouse (Ms), Rat (Rt), Non-human primate (Nhp)
Immunogen: Residues 655-770 from human recombinant protein expressed in bacteria
Ordering Information
Pierce STAT3 Antibody (9D8)
Product #
MA1-13042
Size
100 µL
Price
$330.00
Purchase
Add STAT3 Antibody (9D8) to your cart.
Tested Applications Dilution *
Western Blot (WB) 1:5000
Immunofluorescence (IF) 1:100
Immunohistochemistry (Paraffin) (IHC (P)) 1:1600
Immunoprecipitation (IP) 2 µg
ChIP assay (ChIP) 1-3 ul
* Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own experiment using appropriate negative and positive controls.
Form Information
Form: Liquid
Concentration: 1mg/ml
Purification: Protein A
Storage Buffer: PBS with 1mg/ml BSA
Preservative: 0.05% sodium azide
Storage Conditions: -20° C, Avoid Freeze/Thaw Cycles
Product Specific Information
MA1-13042 detects STAT3 from human, mouse, monkey and rat samples.

MA1-13042 has been successfully used in Western blot, ChIP, immunofluorescence, immunohistochemistry (paraffin), and immunoprecipitation procedures. By Western blot, this antibody detects a 88 kDa protein
corresponding to STAT3. Positive control: HepG2 cells.

The MA1-13042 antigen is a recombinant protein corresponding to residues 655-770 of human STAT3.

Antibodies to this protein (and modification) were previously sold as part of a Thermo Scientific Cellomics High Content Screening Kit. This replacement antibody is now recommended for researchers who need an antibody for high content cell based assays. It has been thoroughly tested and validated for cellular immunofluorescence (IF) applications. Further optimization including the selection of the most appropriate fluorescent Dylight conjugated secondary antibody may have to be performed for your high content assay.
General Information
The protein encoded by this gene is a member of the STAT protein family. In response to cytokines and growth factors, STAT family members are phosphorylated by the receptor associated kinases, and then form homo- or heterodimers that translocate to the cell nucleus where they act as transcription activators. This protein is activated through phosphorylation in response to various cytokines and growth factors including IFNs, EGF, IL5, IL6, HGF, LIF and BMP2. This protein mediates the expression of a variety of genes in response to cell stimuli, and thus plays a key role in many cellular processes such as cell growth and apoptosis. The small GTPase Rac1 has been shown to bind and regulate the activity of this protein. PIAS3 protein is a specific inhibitor of this protein. Three alternatively spliced transcript variants encoding distinct isoforms have been described.
Product Images
  • STAT3 Antibody (MA1-13042) in WB
    Anti-STAT3-Monoclonal-Antibody-9D8-MA1-13042-Western-Blot-1.jpg

    Western Blot with anti-STAT3 Monoclonal Antibody [9D8] (MA1-13042) (Anti-STAT3-Monoclonal-Antibody-9D8-MA1-13042-Western-Blot-1.jpg)

    Western Blot with anti-STAT3 Monoclonal Antibody [9D8] (MA1-13042)

    Western blot analysis of STAT3 was performed by loading 25ug of various whole cell lysates onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% Milk/TBST for at least 1 hour. Membranes were probed with a mouse monoclonal antibody recognizing STAT3 (Product # MA1-13042) at a dilution of 1:5000 overnight at 4°C on a rocking platform. Membranes were washed in TBS-0.1%Tween 20 and probed with a goat anti-mouse-HRP secondary antibody (Product # 32430) at a dilution of 1:20,000 for at least one hour. Membranes were washed and chemiluminescent detection performed using Pierce Super Signal West Dura (Product #34075).
  • STAT3 Antibody (MA1-13042) in WB
    Anti-STAT3-Monoclonal-Antibody-9D8-MA1-13042-Western-Blot-2.jpg

    Western Blot with anti-STAT3 Monoclonal Antibody [9D8] (MA1-13042) (Anti-STAT3-Monoclonal-Antibody-9D8-MA1-13042-Western-Blot-2.jpg)

    Western Blot with anti-STAT3 Monoclonal Antibody [9D8] (MA1-13042)

    Western blot analysis of STAT3 was performed by loading 25ug of HepG2 total lysate onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% Milk/TBST for at least 1 hour. Membranes were probed with a mouse monoclonal antibody recognizing STAT3 (Product # MA1-13042) at a dilution of 1:1000 overnight at 4°C on a rocking platform. Membranes were washed in TBS-0.1%Tween 20 and probed with a goat anti-mouse-HRP secondary antibody (Product # 32430) at a dilution of 1:20,000 for at least one hour. Membranes were washed and chemiluminescent detection performed using Pierce Super Signal West Dura (Product #34075).
  • STAT3 Antibody (MA1-13042) in WB
    Anti-STAT3-Monoclonal-Antibody-9D8-MA1-13042-Western-Blot-3.jpg

    Western Blot with anti-STAT3 Monoclonal Antibody [9D8] (MA1-13042) (Anti-STAT3-Monoclonal-Antibody-9D8-MA1-13042-Western-Blot-3.jpg)

    Western Blot with anti-STAT3 Monoclonal Antibody [9D8] (MA1-13042)

    Western blot analysis of STAT3 was performed by loading 25ug of U2-OS lysate from STAT3 SMART pool siRNA (Product# L-003544-00-0005) transfected or non-targeting control (Product# D-001810-10) transfected U2-OS cells onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% Milk/TBST for at least 1 hour. Membranes were probed with STAT3 (Clone 9D8) mouse monoclonal antibody at a dilution of 1:1000 overnight at 4°C on a rocking platform. Membranes were washed in TBS-0.1%Tween 20 and probed with a goat anti-mouse-HRP secondary antibody (Product # 32430) at a dilution of 1:20,000 for at least one hour. Membranes were washed and chemiluminescent detection performed using Pierce Super Signal West Dura (Product #34075).
  • STAT3 Antibody (MA1-13042) in IF
    Anti-STAT3-Monoclonal-Antibody-9D8-MA1-13042-Immunofluorescent-1.jpg

    Immunofluorescence with anti-STAT3 Monoclonal Antibody [9D8] (MA1-13042) (Anti-STAT3-Monoclonal-Antibody-9D8-MA1-13042-Immunofluorescent-1.jpg)

    Immunofluorescence with anti-STAT3 Monoclonal Antibody [9D8] (MA1-13042)

    Immunofluorescent analysis of STAT3 using anti-STAT3 (9D8) monoclonal antibody (Product# MA1-13042) (shown in green) in HeLa cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were then blocked with 1% Blocker BSA (Product #37525) for 15 minutes at room temperature. Cells were probed with a mouse monoclonal antibody recognizing STAT3 (Product# MA1-13042), at a dilution of 1:100 for at least 1 hour at room temperature. Cells were then washed with PBS and incubated with DyLight 488 goat-anti-mouse secondary antibody (Product# 35503) at a dilution of 1:400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye (Product# 62249). Images were taken on a Thermo Scientific ArrayScan at 20X magnification.
  • STAT3 Antibody (MA1-13042) in IHC
    Anti-STAT3-Monoclonal-Antibody-9D8-MA1-13042-Immunohistochemistry-1.jpg

    Immunohistochemistry with anti-STAT3 Monoclonal Antibody [9D8] (MA1-13042) (Anti-STAT3-Monoclonal-Antibody-9D8-MA1-13042-Immunohistochemistry-1.jpg)

    Immunohistochemistry with anti-STAT3 Monoclonal Antibody [9D8] (MA1-13042)

    Immunohistochemistry was performed on biopsies of normal and cancer tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer for 20 minutes at 95°C. Following antigen retrieval tissues were blocked in 10% normal goat serum (Product #31873) for 20 minutes at room temperature. Tissues were then probed at a dilution of 1:1600 with a mouse monoclonal antibody recognizing STAT3 (Product # MA1-13042) overnight at 4°C in a humidified chamber. Detection was performed using a goat anti-mouse HRP secondary antibody followed by colorimetric detection using DAB substrate. Tissues were counterstained with hematoxylin and prepped for mounting. Images were taken at 400X magnification. Results demonstrate both nuclear and cytoplasmic localization of STAT3.
  • STAT3 Antibody (MA1-13042) in IP
    Anti-STAT3-Monoclonal-Antibody-9D8-MA1-13042-Immunoprecipitation-1.jpg

    Immunoprecipitation with anti-STAT3 Monoclonal Antibody [9D8] (MA1-13042) (Anti-STAT3-Monoclonal-Antibody-9D8-MA1-13042-Immunoprecipitation-1.jpg)

    Immunoprecipitation with anti-STAT3 Monoclonal Antibody [9D8] (MA1-13042)

    Immunoprecipitation of STAT3 was performed on HepG2 cells. The antigen:antibody complex was formed by incubating 750µg whole cell lysate with 2µg of mouse monoclonal antibody recognizing STAT3 (Product # MA1-13042) overnight on a rocking platform at 4°C. The immune-complex was then captured on 50µl Protein A/G Plus Agarose (Product #20423) . Captured immune-complexes were then washed extensively and proteins eluted with 5X Reducing Sample Loading Dye (Product # 39000). Samples were then resolved on a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to PVDF membrane and blocked with 5% Milk/TBS-0.1%Tween for at least 1 hour. Membranes were then probed with a mouse monoclonal antibody recognizing STAT3 (Product # MA1-13042) at a dilution of 1:5000 overnight rotating at 4°C. Membranes were washed in TBST and probed with Clean-blot IP detection reagent (Product #21230) at a dilution of 1:2000 for at least one hour. Membranes were washed and chemiluminescent detection was performed using Pierce Super Signal West Dura (Product #34075).
  • STAT3 Antibody (MA1-13042) in ChIP
    STAT3_Antibody_MA1-13042_ChIP_1_20140428080140.jpg

    ChIP assay with anti-STAT3 Monoclonal Antibody [9D8] (MA1-13042) (STAT3_Antibody_MA1-13042_ChIP_1_20140428080140.jpg)

    ChIP assay with anti-STAT3 Monoclonal Antibody [9D8] (MA1-13042)

    Chromatin immunoprecipitation analysis of STAT3 was performed using cross-linked chromatin from 1x10^6 HTC-IR rat hepatoma cells treated with insulin for 0, 10, and 30 minutes. Immunoprecipitation was performed using a multiplex microplate Matrix ChIP assay (see reference for Matrix ChIP protocol: http://www.ncbi.nlm.nih.gov/pubmed/22098709) with 1.0ul/100ul well volume of a STAT3 monoclonal antibody (Product # MA1-13042). Chromatin aliquots from ~1x10^5 cells were used per ChIP pull-down. Quantitative PCR data were done in quadruplicate using 1ul of eluted DNA in 2ul SYBR real-time PCR reactions containing primers to amplify exon-1 or exon-28 of the LAMC1 gene. PCR calibration curves were generated for each primer pair from a dilution series of sheared total genomic DNA. Quantitation of immunoprecipitated chromatin is presented as signal relative to the total amount of input chromatin. Results represent the mean +/- SEM for three experiments. A schematic representation of the rat LAMC1 locus is shown above the data where boxes represent exons (black boxes = translated regions, white boxes = untranslated regions), the zigzag line represents an intron, and the straight line represents upstream sequence. Regions amplified by LAMC1 primers are represented by black bars. Data courtesy of the Innovators Program.
(This product is for In Vitro experimental use only. Not for resale without express authorization.)
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