Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] - BSA and Azide free (ab236156)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20662-29] to Smad1 (phospho S463 + S465) - BSA and Azide free
  • Suitable for: Dot blot, IP, WB
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] - BSA and Azide free
    See all Smad1 primary antibodies

  • Description

    Rabbit monoclonal [EPR20662-29] 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: Dot blot, IP, WBmore details

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

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

  • Positive control

    • WB: Calyculin and BMP2 treated HeLa cell lysate; Calyculin and BMP2 treated NIH/3T3 cell lysate.

  • General notes

    Ab236156 is the carrier-free version of ab214423. 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

    ab236156 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.

Properties

Images

  • Western blot - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] - BSA and Azide free (ab236156)
    Western blot - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] - BSA and Azide free (ab236156)
    All lanes : Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] (ab214423) at 1/1000 dilution

    Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) grown in serum-free media overnight, whole cell lysate
    Lane 2 : HeLa grown in serum-free media overnight, then treated with 100ng/ml Calyculin A for 15 minutes, Calyculin A was removed, followed by treatment with 100ng/ml BMP2 for 30 minutes, whole cell lysate
    Lane 3 : NIH/3T3 (mouse embryonic fibroblast) grown in serum-free media overnight, whole cell lysate
    Lane 4 : NIH/3T3 grown in serum-free media overnight, then treated with 50ng/ml BMP2 for 30 minutes, whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 52 kDa
    Observed band size: 60 kDa
    why is the actual band size different from the predicted?


    Exposure time: 10 seconds


    Blocking/Dilution: 5% NFDM/TBST

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214423).

  • Immunoprecipitation - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] - BSA and Azide free (ab236156)
    Immunoprecipitation - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] - BSA and Azide free (ab236156)

    Smad1 (phospho S463/S465) was immunoprecipitated from 0.35 mg of NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate with ab214423 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab214423 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1,000 dilution.

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

    Lane 2: ab214423 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 ab214423 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: 30 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214423).

  • Dot Blot - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] - BSA and Azide free (ab236156)
    Dot Blot - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] - BSA and Azide free (ab236156)

    Dot blot analysis of Smad1 (phospho S463/S465) peptide (Lane 1), Smad1 (phospho S463) peptide (Lane 2), Smad1 (phospho S465) peptide (Lane 3), Smad1 non-phospho peptide (Lane 4), Smad5 (phospho S463/S465) peptide (Lane 5), Smad5 (phospho S463) peptide (Lane 6), Smad5 (phospho S465) peptide (Lane 7) and Smad5 non-phospho peptide (Lane 8) using ab214423 at 1/1,000 dilution followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100,000 dilution.

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

    Exposure time: 30 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214423).

  • Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] - BSA and Azide free (ab236156)
    Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] - BSA and Azide free (ab236156)

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