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Overview

  • Product nameAnti-SMAD1/5/9 antibodySee all SMAD1/5/9 primary antibodies ...
  • Description
    Rabbit polyclonal to SMAD1/5/9
  • Tested applicationsICC/IF, WB, IHC-P more details
  • Species reactivity
    Reacts with: Mouse, Rat, Dog, Human, Xenopus laevis, Zebrafish
    Predicted to work with: Cow, Pig, Chimpanzee, Fruit fly (Drosophila melanogaster)
  • Immunogen

    Synthetic peptide: VKGWGAEYHR QDVTSTPCWI EIHLHGPLQW LDKVLTQMGS PHNPISSV, corresponding to C terminal amino acids 417-466 of Human Smad1

  • Positive controlJurkat cell lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab66737 in the following tested applications.

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

Application Notes
ICC/IF ICC/IF: Use a concentration of 1 µg/ml.
WB WB: Use a concentration of 0.625 µg/ml. Detects a band of approximately 62 kDa (predicted molecular weight: 52 kDa). Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
IHC-P IHC-P: Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

  • Alternative names
      AI528653 antibodyBSP 1 antibodyBSP1 antibody
      DKFZp781C1895 antibodyDKFZp781O1323 antibodyDwarfin A antibodyDwf A antibodyDwfA antibodyDwfc antibodyhSMAD1 antibodyhSmad5 antibodyJV 41 antibodyJV4 1 antibodyJV41 antibodyJV5 1 antibodyMAD (mothers against decapentaplegic, Drosophila) homolog 1 antibodyMAD (mothers against decapentaplegic, Drosophila) homolog 9 antibodyMAD homolog 1 antibodyMAD homolog 9 antibodyMAD homolog1 (mothers against decapentaplegic, Drosophila) antibodyMad related protein 1 antibodyMAD, mothers against decapentaplegic homolog 9 antibodyMADH 1 antibodyMadh1 antibodyMADH5 antibodyMADH6 antibodyMADH9 antibodyMadr 1 antibodyMADR1 antibodyMlp1 antibodyMothers against decapentaplegic, drosophila, homolog of, 9 antibodyMothers against DPP homolog 1 antibodyMothers against DPP homolog 9 antibodyMusMLP antibodyOTTHUMP00000178565 antibodyOTTMUSP00000022970 antibodyOTTMUSP00000022971 antibodyOTTMUSP00000022972 antibodyOTTMUSP0000002324 antibodyOTTMUSP00000023428 antibodySma and Mad related protein 1 antibodySma- and Mad-related protein 1 antibodySMAD antibodySMAD family member 5 antibodySMAD mothers against DPP homolog 1 Drosophila antibodySMAD8 antibodySMAD8A antibodySMAD8B antibodySpinal muscular atrophy distal with upper limb predominance antibodyTGF beta signaling protein 1 antibodyTransforming growth factor beta signaling protein 1 antibodyTransforming growth factor-beta signaling protein 1 antibody
    see all

Anti-SMAD1/5/9 antibody images

  • Anti-SMAD1/5/9 antibody (ab66737) at 0.625 µg/ml + Jurkat cell lysate at 10 µg

    Secondary
    HRP conjugated anti-Rabbit IgG at 1/50000 dilution

    Predicted band size : 52 kDa
    Observed band size : 62 kDa (why is the actual band size different from the predicted?)
  • IHC image of ab66737 staining in human heart formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab66737, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    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.

  • ICC/IF image of ab66737 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab66737, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References for Anti-SMAD1/5/9 antibody (ab66737)

This product has been referenced in:
  • Cayli S  et al. Interaction between Smad1 and p97/VCP in rat testis and epididymis during the postnatal development. Reprod Sci 19:190-201 (2012). ICC/IF, WB ; Rat . Read more (PubMed: 22051847) »
  • Liu C  et al. TAK1 promotes BMP4/Smad1 signaling via inhibition of erk MAPK: a new link in the FGF/BMP regulatory network. Differentiation 83:210-9 (2012). WB ; Xenopus laevis . Read more (PubMed: 22387344) »

See all 3 Publications for this product

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Application Western blot
Sample Human Cell lysate - whole cell (Calu6 cell line)
Loading amount 25 µg
Specification Calu6 cell line
Gel Running Conditions Reduced Denaturing (8%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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Verified customer

Submitted Jul 21 2010

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"