Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Smad4 antibody [EP618Y] - BSA and Azide free (ab215968)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP618Y] to Smad4 - BSA and Azide free
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Smad4 antibody [EP618Y] - BSA and Azide free
    See all Smad4 primary antibodies

  • Description

    Rabbit monoclonal [EP618Y] to Smad4 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

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

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

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

  • Positive control

    • WB: NIH/3T3, PC-12, Ramos and SH-SY5Y cell lysates. IHC-P: human lung carcinoma and breast carcinoma tissues.

  • General notes

    Ab215968 is the carrier-free version of ab40759. 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

    ab215968 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    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.

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [EP618Y] - BSA and Azide free (ab215968)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [EP618Y] - BSA and Azide free (ab215968) This image is courtesy of an anonymous Abreview.

    4% PFA-fixed, paraffin-embedded postnatal day 21 mouse ovary tissue stained for Smad4 using ab40759 at 1/500 dilution in immunohistochemical analysis.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40759).

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [EP618Y] - BSA and Azide free (ab215968)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [EP618Y] - BSA and Azide free (ab215968)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue labelling Smad4 with unpurified ab40759 at a 1/100 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40759).

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [EP618Y] - BSA and Azide free (ab215968)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [EP618Y] - BSA and Azide free (ab215968) This image is courtesy of an anonymous Abreview.

    Unpurified ab40759 staining Smad4 in rat femur tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 20 minutes at 22°C; antigen retrieval was by heat mediation in a citrate buffer pH6.0. Samples were incubated with primary antibody (1/200 in blocking buffer) for 2 hours at 20°C. An undiluted HRP-conjugated goat anti-rabbit IgG polyclonal (1/250) was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40759).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [EP618Y] - BSA and Azide free (ab215968)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [EP618Y] - BSA and Azide free (ab215968)

    This IHC data was generated using the same anti-Smad4 antibody clone, EP618Y, in a different buffer formulation (cat# ab40759).

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling Smad4 with purified ab40759 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Western blot - Anti-Smad4 antibody [EP618Y] - BSA and Azide free (ab215968)
    Western blot - Anti-Smad4 antibody [EP618Y] - BSA and Azide free (ab215968)

    This WB data was generated using the same anti-Smad4 antibody clone, EP618Y, in a different buffer formulation (cat# ab40759).

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: SMAD4 knockout  HAP1 whole cell lysate (20 µg)
    Lane 3: HepG2 whole cell lysate (20 µg)
    Lane 4: Jurkat whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab40759 observed at 60 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab40759 was shown to specifically react with SMAD4 in wild type HAP1 cells. No band was observed when SMAD4 knockout HAP1 samples were used. Wild-type and SMAD4 knockout samples were subjected to SDS-PAGE. Ab40759 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1000 dilution and 1/20000 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 1/20000 dilution for 1 hour at room temperature before imaging.

  • Anti-Smad4 antibody [EP618Y] - BSA and Azide free (ab215968)
    Anti-Smad4 antibody [EP618Y] - BSA and Azide free (ab215968)

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