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RecombinantRabMAb

Recombinant Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)

Submit a review Submit a question References (1)
Immunoprecipitation - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)
  • Flow Cytometry (Intracellular) - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)
  • Immunoprecipitation - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)
  • ChIP - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)
  • Immunocytochemistry/ Immunofluorescence - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)
  • Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19557] to Smad2 + Smad3 - BSA and Azide free
  • Suitable for: Flow Cyt (Intra), ChIP, IP, ICC/IF, WB
  • Reacts with: Human, Recombinant fragment

Conjugates logo Related conjugates and formulations

Unconjugated

Overview

  • Product name

    Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free
    See all Smad2 + Smad3 primary antibodies

  • Description

    Rabbit monoclonal [EPR19557] to Smad2 + Smad3 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt (Intra), ChIP, IP, ICC/IF, WBmore details

  • Species reactivity

    Reacts with: Human, Recombinant fragment

  • Immunogen

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

  • Positive control

    • ICC/IF: HeLa cells.

  • General notes

    ab236030 is the carrier-free version of ab207447.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    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.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. 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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab236030 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
Flow Cyt (Intra)
Use at an assay dependent concentration.
ChIP
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 58-62 kDa (predicted molecular weight: 52 kDa).
Notes
Flow Cyt (Intra)
Use at an assay dependent concentration.
ChIP
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 58-62 kDa (predicted molecular weight: 52 kDa).

Target

  • Relevance

    SMAD is a family of proteins similar to the gene products of the Drosophila gene 'mothers against decapentaplegic' (Mad) and the C. elegans gene Sma. SMAD proteins are signal transducers and transcriptional modulators that mediate multiple signaling pathways. They mediate the signal of the transforming growth factor (TGF)-beta, and thus regulate multiple cellular processes, such as cell proliferation, apoptosis, and differentiation.

  • Cellular localization

    Cytoplasm. Nucleus. Note: Cytoplasmic in the absence of ligand. Migrates to the nucleus when complexed with SMAD4.

  • Database links

    • Alternative names

      • hMAD 2 antibody
      • hMAD 3 antibody
      • hSMAD2 antibody
      • hSMAD3 antibody
      • Mad related protein 2 antibody
      • MADH2 antibody
      • MADH3 antibody
      • MADR2 antibody
      • Mothers against DPP homolog 2 antibody
      • Mothers against DPP homolog 3 antibody
      • Sma and Mad related protein 2 antibody
      • SMA and MAD related protein 3 antibody
      • SMAD 2 antibody
      • SMAD 3 antibody
      • SMAD family member 2 antibody
      • SMAD family member 3 antibody
      • smad2+3 antibody
      • smad2/3 antibody
      see all

    Images

    • Immunoprecipitation - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)
      Immunoprecipitation - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)

      ab207447 Immunoprecipitating Smad2 + Smad3 in human HeLa whole cell lysate . 2µg of capture antibody in 0.35mg lysate was used. 10µg of cell lysate was incubated with primary antibody at a dilution of 1/1000 and VeriBlot for IP Detection Reagent (HRP) ab131366 was used for detection at a dilution of 1/1000.

      Lane 1: HeLa (human cervix adenocarcinoma) whole cell lysate
      Lane 2: HeLa (human cervix adenocarcinoma) whole cell lysate
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab207447 in HeLa (human cervix adenocarcinoma) whole cell lysate

      Blocking buffer: 5% NFDM/TBST

      Diluting buffer: 5% NFDM/TBST

      Smad2 and Smad3 can be resolved using a lower percentage gel.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207447).

    • Flow Cytometry (Intracellular) - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)
      Flow Cytometry (Intracellular) - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)

      Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa  (Human epithelial cell line from cervix adenocarcinoma) cells labeling Smad2 and Smad3 with ab207447 at 1/500 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/500 dilution 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 (ab207447). 

       

       

    • Immunoprecipitation - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)
      Immunoprecipitation - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)

      Smad2 + Smad3 were immunoprecipitated from 1mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab207447 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab207447 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

      Lane 1: HeLa whole cell lysate 10µg (Input).

      Lane 2: ab207447 IP in HeLa whole cell lysate.

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab207447 in HeLa whole cell lysate.

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

      Exposure time: 5 seconds.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207447).

    • ChIP - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)
      ChIP - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)

      Chromatin was prepared from HaCaT (Human keratinocyte cell line) cells treated with 7ng/ml TGF-β for 1h and non-treated according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab207447 (blue), and 20µl of Anti rabit IgG sepharose beads. 2μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

      The ChIP condition is designed against Smad2 refer to PMID: 18955504.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207447).

    • Immunocytochemistry/ Immunofluorescence - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)
      Immunocytochemistry/ Immunofluorescence - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Smad2 + Smad3 with ab207447 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing signal translocation from cytoplasm to nucleus after TGF-beta (10ng/ml, 1h) treatment in HeLa cells. PMID: 9006934. The nuclear counter stain is DAPI (blue).

      Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

      The negative controls are as follows:

      -ve control 1: ab207447 at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary at 1/1000 dilution.

      -ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary 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 (ab207447).

    • Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)
      Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (1)

    Publishing research using ab236030? Please let us know so that we can cite the reference in this datasheet.

    ab236030 has been referenced in 1 publication.

    • Bai Y  et al. Mesenchymal Stem Cells Reverse Diabetic Nephropathy Disease via Lipoxin A4 by Targeting Transforming Growth Factor ß (TGF-ß)/smad Pathway and Pro-Inflammatory Cytokines. Med Sci Monit 25:3069-3076 (2019). PubMed: 31023998

    Customer reviews and Q&As

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