Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-STAT3 antibody [EPR361] - BSA and Azide free (ab171360)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR361] to STAT3 - BSA and Azide free
  • Suitable for: IHC-P, ICC/IF, WB, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-STAT3 antibody [EPR361] - BSA and Azide free
    See all STAT3 primary antibodies

  • Description

    Rabbit monoclonal [EPR361] to STAT3 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, ICC/IF, WB, Flow Cyt (Intra)more details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HAP1, HEK293 and HeLa cell lysates. IHC-P: Human pancreas tissue. ICC/IF: A459 and HeLa cells. Flow Cyt (intra): HeLa cells.

  • General notes

    ab171360 is the carrier-free version of ab109085.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

Images

  • Western blot - Anti-STAT3 antibody [EPR361] - BSA and Azide free (ab171360)
    Western blot - Anti-STAT3 antibody [EPR361] - BSA and Azide free (ab171360)
    All lanes : Anti-STAT3 antibody [EPR361] (ab109085) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : STAT3 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 88 kDa
    Observed band size: 92 kDa
    why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab109085).

      Lanes 1- 2: Merged signal (red and green). Green - ab109085 observed at 92 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab109085 was shown to react with STAT3 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255436 (knockout cell lysate ab263797) was used. Wild-type HeLa and STAT3 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109085 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-STAT3 antibody [EPR361] - BSA and Azide free (ab171360)
    Immunocytochemistry/ Immunofluorescence - Anti-STAT3 antibody [EPR361] - BSA and Azide free (ab171360)

    Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labeling STAT3 with purified ab109085 at 1/250 dilution (5 µg/ml). Cells were fixed with 4% PFA and permeabilized with 0.1% tritonX-100. ab150077 Goat anti rabbit IgG (Alexa Fluor® 488) at 1/1000 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109085).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 antibody [EPR361] - BSA and Azide free (ab171360)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 antibody [EPR361] - BSA and Azide free (ab171360)

    Immunohistochemical staining of paraffin embedded human pancreas with purified ab109085 at a working dilution of 1/100. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109085).

  • Western blot - Anti-STAT3 antibody [EPR361] - BSA and Azide free (ab171360)
    Western blot - Anti-STAT3 antibody [EPR361] - BSA and Azide free (ab171360)
    All lanes : Anti-STAT3 antibody [EPR361] (ab109085) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : STAT3 knockout HAP1 whole cell lysate
    Lane 3 : HeLa whole cell lysate
    Lane 4 : HEK293 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 88 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab109085).

    Lanes 1 - 4: Merged signal (red and green). Green - ab109085 observed at 92 kDa. Red - loading control, ab8245, observed at 37 kDa.

    Ab109085 was shown to specifically react with STAT3 in wild-type cells as signal was lost in STAT3 knockout HAP1 cells. Wild-type and STAT3 knockout samples were subjected to SDS-PAGE. Ab109085 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-STAT3 antibody [EPR361] - BSA and Azide free (ab171360)
    Immunocytochemistry/ Immunofluorescence - Anti-STAT3 antibody [EPR361] - BSA and Azide free (ab171360)

    Immunofluorescence staining of A459 cells with purified ab109085 at a working dilution of 1/150, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 555 goat anti rabbit (ab150078), used at a dilution of 1/500. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative control is shown in bottom right hand panel - for the negative control, purified ab109085 was used at a dilution of 1/200 followed by an Alexa Fluor® 488 goat anti-mouse antibody at a dilution of 1/500.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109085).

  • Flow Cytometry - Anti-STAT3 antibody [EPR361] - BSA and Azide free (ab171360)
    Flow Cytometry - Anti-STAT3 antibody [EPR361] - BSA and Azide free (ab171360)

    Flow cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling STAT3 (red) with ab109085 at a 1/300 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109085).

  • Anti-STAT3 antibody [EPR361] - BSA and Azide free (ab171360)
    Anti-STAT3 antibody [EPR361] - BSA and Azide free (ab171360)

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