Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-STAT1 antibody [EPR23049-111] - ChIP Grade (ab239360)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23049-111] to STAT1 - ChIP Grade
  • Suitable for: Flow Cyt, ICC/IF, ChIP, WB, IP
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-STAT1 antibody [EPR23049-111] - ChIP Grade
    See all STAT1 primary antibodies

  • Description

    Rabbit monoclonal [EPR23049-111] to STAT1 - ChIP Grade

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ChIP
    Human
    Flow Cyt
    Mouse
    ICC/IF
    Mouse
    Human
    IP
    Mouse
    WB
    Mouse
    Human
    See all applications and species data

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human lymph node, RAW264.7, HeLa, MCF7, Mouse lung, Mouse breast and NIH/3T3 lysates. ICC/IF: HeLa and RAW 264.7 cells. Flow Cyt: RAW264.7 cells. IP: RAW264.7 cells.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Images

  • ChIP - Anti-STAT1 antibody [EPR23049-111] (ab239360)
    ChIP - Anti-STAT1 antibody [EPR23049-111] (ab239360)

    Chromatin was prepared from HT-1080 cells treated with IFN-γ (50ng/ml 30min) according to the Abcam Dual-X-ChIP protocol*. Cells were fixed with 1.5 mM EGS for 30mins and then formaldehyde for 10min.

    The  ChIP was performed with 25 µg of chromatin, 5 µg of  ab239360 (red), or 5 µg of rabbit normal IgG ab172730 (gray) and 20 µl of Protein A/G sepharose beads.  The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci).

    Primers and probes are commercial primers from Cell Signaling Technology (Cat. No.: #5148)

    *http://www.abcam.com/resources?keywords=X%20ChIP%20protocol

  • Western blot - Anti-STAT1 antibody [EPR23049-111] (ab239360)
    Western blot - Anti-STAT1 antibody [EPR23049-111] (ab239360)
    All lanes : Anti-STAT1 antibody [EPR23049-111] - ChIP Grade (ab239360) at 1/1000 dilution

    Lane 1 : Human lymph node tissue lysate
    Lane 2 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate
    Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
    Lane 4 : MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate
    Lane 5 : Mouse lung tissue lysate
    Lane 6 : Mouse breast tissue lysate
    Lane 7 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 87 kDa
    Observed band size: 88 kDa
    why is the actual band size different from the predicted?



    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    Lanes 5-7 were developed using a higher sensitivity ECL substrate. 

    Exposure time: Lanes 1-4: 3 minutes; Lane 5-7: 48 seconds.

  • Immunocytochemistry/ Immunofluorescence - Anti-STAT1 antibody [EPR23049-111] (ab239360)
    Immunocytochemistry/ Immunofluorescence - Anti-STAT1 antibody [EPR23049-111] (ab239360)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling STAT1 with ab239360 at 1/50 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1\1000 dilution (Green). Confocal image showing the signal translocate to nuclear in HeLa cells treated with IFN-γ (10ng/ml) for 30 mins. Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

  • Flow Cytometry - Anti-STAT1 antibody [EPR23049-111] (ab239360)
    Flow Cytometry - Anti-STAT1 antibody [EPR23049-111] (ab239360)

    Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling STAT1 with ab239360 at 1/600 (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 was used as the secondary antibody.

  • Immunoprecipitation - Anti-STAT1 antibody [EPR23049-111] (ab239360)
    Immunoprecipitation - Anti-STAT1 antibody [EPR23049-111] (ab239360)

    STAT1 was immunoprecipitated from 0.35mg RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate with ab239360 at     1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab239360 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

    Lane 1: RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate (10ug)

    Lane 2: ab239360 IP in RAW264.7 whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab239360 in RAW264.7 whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 75 seconds.

     

  • Immunocytochemistry/ Immunofluorescence - Anti-STAT1 antibody [EPR23049-111] (ab239360)
    Immunocytochemistry/ Immunofluorescence - Anti-STAT1 antibody [EPR23049-111] (ab239360)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling STAT1 with ab239360 at 1/50 dilution, followed byab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing the signal translocate to nuclear in RAW 264.7 cells treated with IFN-γ (10ng/ml) for 30 mins. Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

  • Anti-STAT1 antibody [EPR23049-111] - ChIP Grade (ab239360)
    Anti-STAT1 antibody [EPR23049-111] - ChIP Grade (ab239360)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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