Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-STAT5b antibody [EPR16671] - BSA and Azide free (ab240211)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR16671] to STAT5b - BSA and Azide free
  • Suitable for: WB, ICC/IF, IP, IHC-P, Flow Cyt, ChIP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-STAT5b antibody [EPR16671] - BSA and Azide free
    See all STAT5b primary antibodies

  • Description

    Rabbit monoclonal [EPR16671] to STAT5b - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ChIP
    Human
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Rat
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

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

  • General notes

    Ab240211 is the carrier-free version of ab178941. 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

    ab240211 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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Images

  • Western blot - Anti-STAT5b antibody [EPR16671] - BSA and Azide free (ab240211)
    Western blot - Anti-STAT5b antibody [EPR16671] - BSA and Azide free (ab240211)
    All lanes : Anti-STAT5b antibody [EPR16671] (ab178941) at 1/20000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : STAT5B knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 90 kDa
    Observed band size: 90 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab178941).

    Lanes 1-2: Merged signal (red and green). Green - ab178941 observed at 90 kDa. Red - loading control ab8245 observed at 37 kDa.

     ab178941 Anti-STAT5b antibody [EPR16671] was shown to specifically react with STAT5b in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab266006 (knockout cell lysate ab257710) was used. Wild-type and STAT5b knockout samples were subjected to SDS-PAGE. ab178941 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 20000 and 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.

     

     

  • Immunoprecipitation - Anti-STAT5b antibody [EPR16671] - BSA and Azide free (ab240211)
    Immunoprecipitation - Anti-STAT5b antibody [EPR16671] - BSA and Azide free (ab240211)

    Immunoprecipitation of K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell extract using ab178941 at 1/40 dilution. Western blot detection of STAT5b utilised ab178941 at 1/2000 dilution and Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated secondary antibody at 1/1000 dilution. The blocking and dilution buffer was 5% NFDM/TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178941).

  • ChIP - Anti-STAT5b antibody [EPR16671] - BSA and Azide free (ab240211)
    ChIP - Anti-STAT5b antibody [EPR16671] - BSA and Azide free (ab240211)

    Chromatin was prepared from T-47D (starved overnight) treated with Prolactin(10nM 30min) and T-47D (starved overnight) non-treated cells 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 ab178941 (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 (Sybr green approach).
    Primers are from PMID: 15686596.
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178941).

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

     

  • Immunocytochemistry/ Immunofluorescence - Anti-STAT5b antibody [EPR16671] - BSA and Azide free (ab240211)
    Immunocytochemistry/ Immunofluorescence - Anti-STAT5b antibody [EPR16671] - BSA and Azide free (ab240211)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling STAT5b with ab178941 at 1/100 dilution. The cells were permeabilised with 0.1% Triton X-100. Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/200 dilution was used as the secondary antibody (green). Nuclear and cytoplasm staining is detected. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (Tubulin mouse mAb) at 1/500 and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/400 dilution (red). 

     

    The negative controls are as follows;

    1. ab178941 at 1/100 dilution followed by Goat anti mouse IgG (Alexa Fluor®594) at 1/400 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by Goat anti rabbit IgG (Alexa Fluor®488) ar 1/200 dilution.

     

     

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178941).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5b antibody [EPR16671] - BSA and Azide free (ab240211)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5b antibody [EPR16671] - BSA and Azide free (ab240211)

    Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling STAT5b with ab178941 at 1/500 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Nucleus staining on lymphocytes of Human spleen is detected. The negative control utilised PBS instead of primary antibody. Counter stained with Hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178941).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Flow Cytometry - Anti-STAT5b antibody [EPR16671] - BSA and Azide free (ab240211)
    Flow Cytometry - Anti-STAT5b antibody [EPR16671] - BSA and Azide free (ab240211)

    ab178941 staining STAT5b in the human cell line HeLa (human cervix adenocarcinoma) by flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/40. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

    Isoytype control: Rabbit monoclonal IgG (Black)

    Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178941).

  • Immunoprecipitation - Anti-STAT5b antibody [EPR16671] - BSA and Azide free (ab240211)
    Immunoprecipitation - Anti-STAT5b antibody [EPR16671] - BSA and Azide free (ab240211)

    Cross Immunoprecipitation of K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell extract showing no cross reactivity with STAT5a. Protein captured by anti-STAT5a antibody (ab32042) was detected by the same antibody in WB (image 1) but not by anti-STAT5b, ab178941 (image 2).

     

    For WB detection, ab178941 was used at a 1/2000 dilution and Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated secondary antibody at a 1/1000 dilution. The blocking and dilution buffer was 5% NFDM/TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178941).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5b antibody [EPR16671] - BSA and Azide free (ab240211)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5b antibody [EPR16671] - BSA and Azide free (ab240211)

    Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling STAT5b with ab178941 at 1/500 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Nucleus staining on lymphocytes of Mouse spleen is detected. The negative control utilised PBS instead of primary antibody. Counter stained with Hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178941).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5b antibody [EPR16671] - BSA and Azide free (ab240211)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5b antibody [EPR16671] - BSA and Azide free (ab240211)

    Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling STAT5b with ab178941 at 1/500 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Nucleus staining on lymphocytes and weak nucleus staining on gland epithelium of colon is detected. The negative control utilised PBS insead of primary antibody and the slide is counter stained with Hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178941).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Flow Cytometry - Anti-STAT5b antibody [EPR16671] - BSA and Azide free (ab240211)
    Flow Cytometry - Anti-STAT5b antibody [EPR16671] - BSA and Azide free (ab240211)

    Flow cytometry analysis of 2% paraformaldehyde K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling STAT5b with ab178941 at 1:60 dilution (red line). Secondary antibody used is a goat anti rabbit IgG (FITC) at 1:150 dilution. The isotype control is rabbit monoclonal IgG (black line).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178941).

  • Anti-STAT5b antibody [EPR16671] - BSA and Azide free (ab240211)
    Anti-STAT5b antibody [EPR16671] - BSA and Azide free (ab240211)

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