Astana Biomed Group, an authorized Abcam distributor in Central Asia

HRP Anti-Smad4 antibody [EP618Y] (ab195554)

Key features and details

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

Overview

  • Product name

    HRP Anti-Smad4 antibody [EP618Y]
    See all Smad4 primary antibodies

  • Description

    HRP Rabbit monoclonal [EP618Y] to Smad4

  • Host species

    Rabbit

  • Conjugation

    HRP

  • Tested applications

    Suitable for: IHC-P, WBmore details

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat

  • Immunogen

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

  • Positive control

    • WB: SH SY5Y whole cell lysate. IHC-P: normal human colon tissue.

  • General notes

     

     

    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 - HRP Anti-Smad4 antibody [EP618Y] (ab195554)
    Western blot - HRP Anti-Smad4 antibody [EP618Y] (ab195554)
    All lanes : HRP Anti-Smad4 antibody [EP618Y] (ab195554) at 1/5000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : SMAD4 knockout HAP1 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 65 kDa
    Observed band size: 65 kDa


    Exposure time: 12 minutes


    ab195554 was shown to recognize Smad4 in wild-type HAP1 cells as signal was lost at the expected MW in SMAD4 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and SMAD4 knockout samples were subjected to SDS-PAGE. Ab195554 and ab184095 (Mouse monoclonal [mAbcam 9484] to GAPDH - Loading Control (Alexa Fluor® 680) loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 dilution respectively. The loading control was imaged using the Licor Odyssey CLx prior to blots being developed with ECL technique.

  • Western blot - HRP Anti-Smad4 antibody [EP618Y] (ab195554)
    Western blot - HRP Anti-Smad4 antibody [EP618Y] (ab195554)
    HRP Anti-Smad4 antibody [EP618Y] (ab195554) at 1/5000 dilution + SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate at 10 µg

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 65 kDa
    Observed band size: 65 kDa


    Exposure time: 20 minutes


    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab195554 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HRP Anti-Smad4 antibody [EP618Y] (ab195554)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HRP Anti-Smad4 antibody [EP618Y] (ab195554)

    IHC image of Smad4 staining in a section of formalin-fixed paraffin-embedded normal human colon tissue*, performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab195554 at 1/100 dilution, for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • HRP Anti-Smad4 antibody [EP618Y] (ab195554)
    HRP Anti-Smad4 antibody [EP618Y] (ab195554)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Alternative products to HRP Anti-Smad4 antibody [EP618Y] (ab195554)

© 2021 Astana Biomed Group LLP. All rights reserved.