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Anti-STAT3 antibody [E121-21] - BSA and Azide free (ab171361)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [E121-21] to STAT3 - BSA and Azide free
  • Suitable for: ChIP, Flow Cyt, ICC/IF, WB, IP, IHC-P
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-STAT3 antibody [E121-21] - BSA and Azide free
    See all STAT3 primary antibodies

  • Description

    Rabbit monoclonal [E121-21] to STAT3 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ChIP, Flow Cyt, ICC/IF, WB, IP, IHC-Pmore details

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Horse, Cow

  • Immunogen

    Synthetic peptide corresponding to Human STAT3. A synthetic peptide corresponding to residues surrounding Ser727 of human Stat3.

  • General notes

    Ab171361 is the carrier-free version of ab32500. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

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

    ab171361 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Images

  • Western blot - Anti-STAT3 antibody [E121-21] - BSA and Azide free (ab171361)
    Western blot - Anti-STAT3 antibody [E121-21] - BSA and Azide free (ab171361)

    This WB data was generated using the same anti-STAT3 antibody clone, E121-21, in a different buffer formulation (cat# ab32500).

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: STAT3 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lane 4: HEK293 whole cell lysate (20 µg)

     

    Lanes 1 - 4: Merged signal (red and green). Green - ab32500 observed at 92 kDa. Red - loading control, ab8245, observed at 37 kDa.

     

    Ab32500 detected the expected band for STAT3 in wild-type cells along with additional cross-reactive bands. The band was not seen in STAT3 knockout HAP1 cells. Wild-type and STAT3 knockout samples were subjected to SDS-PAGE. Ab32500 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 [E121-21] - BSA and Azide free (ab171361)
    Immunocytochemistry/ Immunofluorescence - Anti-STAT3 antibody [E121-21] - BSA and Azide free (ab171361)

    ab32500 staining STAT3 in HeLa cells. The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab32500 at 1/250 dilution (shown in green) and ab195889, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor® 594), at 2µg/ml (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32500).

  • Flow Cytometry - Anti-STAT3 antibody [E121-21] - BSA and Azide free (ab171361)
    Flow Cytometry - Anti-STAT3 antibody [E121-21] - BSA and Azide free (ab171361)

    Overlay histogram showing HeLa cells stained with ab32500 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween 20 for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32500, 1/100) for 30 min at 22ºC. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150081) at 1/2000 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (ab172730, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32500).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 antibody [E121-21] - BSA and Azide free (ab171361)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 antibody [E121-21] - BSA and Azide free (ab171361)

    This IHC data was generated using the same anti-STAT3 antibody clone, E121-21, in a different buffer formulation (cat# ab32500).

    Immunohistochemical analysis of paraffin-embedded lung squamous carcinoma using ab32500 at a 1/50 dilution.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • ChIP - Anti-STAT3 antibody [E121-21] - BSA and Azide free (ab171361)
    ChIP - Anti-STAT3 antibody [E121-21] - BSA and Azide free (ab171361)

    Chromatin was prepared from HepG2 + IL-6 (100 ng/ml 30 minutes) cells according to the Abcam X-ChIP protocol*. Cells were fixed with formaldehyde for 10 minutes.
    The ChIP was performed with 25 µg of chromatin, 2 µg of ab32500 (red), and 20 µl of Protein A/G sepharose beads. 2 µg of rabbit normal IgG was added to the beads control (grey). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
    Primers and probes are located in the first kb of the transcribed region.
    *http///www.abcam.com/resources?keywords=X%20ChIP%20protocol
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32500).

  • Immunoprecipitation - Anti-STAT3 antibody [E121-21] - BSA and Azide free (ab171361)
    Immunoprecipitation - Anti-STAT3 antibody [E121-21] - BSA and Azide free (ab171361)

    ab32500 (purified) at 1/500 dilution (2.594 µg/ml) immunoprecipitating STAT3 in HeLa whole cell lysate.

    Lane 1 (input): HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
    Lane 2 (+): ab32500 & HeLa whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32500 in HeLa whole cell lysate

    For western blotting, VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/1000 dilution.

    Blocking and diluting buffer: 5% NFDM /TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32500).

  • Anti-STAT3 antibody [E121-21] - BSA and Azide free (ab171361)
    Anti-STAT3 antibody [E121-21] - BSA and Azide free (ab171361)

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