Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-20] - BSA and Azide free (ab228457)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20662-20] to Smad1 (phospho S463 + S465) - BSA and Azide free
  • Suitable for: IP, WB, ICC/IF, Dot blot
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-20] - BSA and Azide free
    See all Smad1 primary antibodies

  • Description

    Rabbit monoclonal [EPR20662-20] to Smad1 (phospho S463 + S465) - BSA and Azide free

  • Host species

    Rabbit

  • Specificity

    Based on sequence homology this antibody also reacts with Smad5 (phospho S463/S465) and Smad9 (phospho S465/S467).

  • Tested applications

    Suitable for: IP, WB, ICC/IF, Dot blotmore details

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

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

  • Positive control

    • ICC/IF: NIH3T3 cells FBS-deprived overnight before treatment with 50 ng/ml hBMP2 for 30min.

  • General notes

    Ab228457 is the carrier-free version of ab226821. 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

    ab228457 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

  • Dot Blot - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-20] - BSA and Azide free (ab228457)
    Dot Blot - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-20] - BSA and Azide free (ab228457)

    Dot blot analysis of Smad1 (phospho S463 + S465) labeled with ab226821 at 1/1000 dilution.

    Lane 1: Smad1 (phospho S463/S465) peptide;

    Lane 2: Smad1 (phospho S463) peptide;

    Lane 3: Smad1 (phospho S465) peptide;

    Lane 4: Smad1 peptide (not phosphorylated);

    Lane 5: Smad5 (phospho S463/S465) peptide;

    Lane 6: Smad5 (phospho S463) peptide;

    Lane 7: Smad5 (phospho S465) peptide;

    Lane 9: Smad5 peptide (not phosphorylated).

    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution was used as secondary antibody.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: 3 minutes.

    Based on sequence homology, this antibody cross-reacts with Smad5 (phospho S463/S465) and Smad9 (phospho S465/S467).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab226821).

  • Immunoprecipitation - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-20] - BSA and Azide free (ab228457)
    Immunoprecipitation - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-20] - BSA and Azide free (ab228457)

    Smad 1 (phospho S463 + S465) was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryo fibroblast cell line) grown in serum-free media overnight, then treated with 50 ng/ml BMP2 for 30 minutes, whole cell lysate with ab226821 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab226821 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

    Lane 1: NIH/3T3 grown in serum-free media overnight, then treated with 50 ng/ml BMP2 for 30 minutes, whole cell lysate 10 μg (Input).

    Lane 2: ab226821 IP in NIH/3T3 grown in serum-free media overnight, then treated with 50 ng/ml BMP2 for 30 minutes, whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab226821 in NIH/3T3 grown in serum-free media overnight, then treated with 50 ng/ml BMP2 for 30 minutes, whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 10 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab226821).

  • Immunocytochemistry/ Immunofluorescence - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-20] - BSA and Azide free (ab228457)
    Immunocytochemistry/ Immunofluorescence - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-20] - BSA and Azide free (ab228457)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryo fibroblast cell line) cells labeling Smad1 (phospho S463 + S465) with ab226821 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Nuclear staining in hBMP2-treated NIH/3T3 cells. Cells were FBS-deprived overnight before treatment with 50 ng/ml hBMP2 for 30 minutes.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab226821).

  • Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-20] - BSA and Azide free (ab228457)
    Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-20] - BSA and Azide free (ab228457)

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