Recombinant
RabMAb

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

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 within Human Smad1 aa 450 to the C-terminus. The exact sequence is proprietary.
    Database link: Q15797

  • Positive control

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

    The formulation and the concentration of this product is compatible for metal-conjugation for mass cytometry (CyTOF®).

    ab236156 is a PBS-only buffer format of ab214423. Please refer to ab214423 for recommended dilutions, protocols, and image data. 

     

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab236156 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Dot blot Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 60 kDa (predicted molecular weight: 52 kDa).

Target

  • Function

    Transcriptional modulator activated by BMP (bone morphogenetic proteins) type 1 receptor kinase. SMAD1 is a receptor-regulated SMAD (R-SMAD). SMAD1/OAZ1/PSMB4 complex mediates the degradation of the CREBBP/EP300 repressor SNIP1.
  • Tissue specificity

    Ubiquitous. Highest expression seen in the heart and skeletal muscle.
  • Sequence similarities

    Belongs to the dwarfin/SMAD family.
    Contains 1 MH1 (MAD homology 1) domain.
    Contains 1 MH2 (MAD homology 2) domain.
  • Post-translational
    modifications

    Phosphorylated on serine by BMP type 1 receptor kinase.
    Ubiquitin-mediated proteolysis by SMAD-specific E3 ubiquitin ligase SMURF1.
  • Cellular localization

    Cytoplasm. Nucleus. Cytoplasmic in the absence of ligand. Migrates to the nucleus when complexed with SMAD4. Co-localizes with LEMD3 at the nucleus inner membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • BSP-1 antibody
    • BSP1 antibody
    • HsMAD1 antibody
    • JV4-1 antibody
    • JV41 antibody
    • MAD homolog 1 antibody
    • MAD mothers against decapentaplegic homolog 1 antibody
    • Mad related protein 1 antibody
    • Mad-related protein 1 antibody
    • MADH1 antibody
    • MADR1 antibody
    • Mothers against decapentaplegic homolog 1 antibody
    • Mothers against DPP homolog 1 antibody
    • SMA- AND MAD-RELATED PROTEIN 1 antibody
    • SMAD 1 antibody
    • SMAD family member 1 antibody
    • SMAD mothers against DPP homolog 1 antibody
    • Smad1 antibody
    • SMAD1_HUMAN antibody
    • TGF beta signaling protein 1 antibody
    • Transforming growth factor-beta-signaling protein 1 antibody
    see all

Images

  • 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).

  • 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 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).

References

ab236156 has not yet been referenced specifically in any publications.

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