Overview

  • Product name

  • Description

    Rabbit polyclonal to SMAD1/5/9
  • Host species

    Rabbit
  • Specificity

    ab80255 detects endogenous levels of total SMAD1/5/9 protein.
  • Tested applications

    Suitable for: IHC-FoFr, WB, ELISAmore details
  • Species reactivity

    Reacts with: Rat, Human
    Predicted to work with: Mouse
  • Immunogen

    Synthetic peptide derived from N terminal of human SMAD1/5/9

  • Positive control

    • HeLa cell extracts (WB), P7 rat cortex (IHC)

Properties

Applications

Our Abpromise guarantee covers the use of ab80255 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
IHC-FoFr 1/500.
WB 1/500 - 1/1000. Detects a band of approximately 52 kDa (predicted molecular weight: 52 kDa).
ELISA 1/10000.

Target

  • Relevance

    SMADs are members of the MAD-related family of molecules. MAD-related proteins are a family of intracellular proteins that are essential components in the signaling pathways of the serine/threonine kinase receptors of the transforming growth factor beta superfamily. SMADs can be divided into receptor-regulated SMADs, common-mediator SMAD and inhibitory SMADs. SMAD1, SMAD5 and SMAD9 have high degrees of homology. SMAD8 and SMAD9 are used as alternate names for the same protein. Human SMAD1 is a 465 amino acid protein, human SMAD5 is a 465 amino acid and human SMAD9 is a 430 amino acid protein.
  • Cellular localization

    Cytoplasmic and Nuclear
  • Database links

  • Alternative names

    • AI528653 antibody
    • BSP1 antibody
    • DKFZp781C1895 antibody
    • DKFZp781O1323 antibody
    • Dwarfin A antibody
    • Dwf A antibody
    • Dwfc antibody
    • hSMAD1 antibody
    • hSmad5 antibody
    • JV4 1 antibody
    • JV41 antibody
    • JV5 1 antibody
    • MAD (mothers against decapentaplegic, Drosophila) homolog 1 antibody
    • MAD (mothers against decapentaplegic, Drosophila) homolog 9 antibody
    • MAD homolog 1 antibody
    • MAD homolog 9 antibody
    • MAD homolog1 (mothers against decapentaplegic, Drosophila) antibody
    • Mad related protein 1 antibody
    • MAD, mothers against decapentaplegic homolog 9 antibody
    • Madh1 antibody
    • MADH5 antibody
    • MADH6 antibody
    • MADH9 antibody
    • MADR1 antibody
    • Mlp1 antibody
    • Mothers against decapentaplegic, drosophila, homolog of, 9 antibody
    • Mothers against DPP homolog 1 antibody
    • Mothers against DPP homolog 9 antibody
    • MusMLP antibody
    • OTTHUMP00000178565 antibody
    • OTTMUSP00000022970 antibody
    • OTTMUSP00000022971 antibody
    • OTTMUSP00000022972 antibody
    • OTTMUSP0000002324 antibody
    • OTTMUSP00000023428 antibody
    • Sma- and Mad-related protein 1 antibody
    • SMAD antibody
    • SMAD family member 5 antibody
    • SMAD8 antibody
    • SMAD8A antibody
    • SMAD8B antibody
    • TGF beta signaling protein 1 antibody
    • Transforming growth factor-beta signaling protein 1 antibody
    see all

Images

  • All lanes : Anti-SMAD1/5/9 antibody (ab80255) at 1/500 dilution

    Lane 1 : HeLa cell extracts
    Lane 2 : HeLa cell extracts with immunising peptide at 10 µg

    Lysates/proteins at 30 µg per lane.

    Predicted band size: 52 kDa
    Observed band size: 52 kDa

  • Immunohistomchemical detection of SMAD1/5/9 using antibody ab80255 on perfusion fixed P7 rat cortex sections. The animals were perfused with 4% PFA. The sections were incubated in 10% normal donkey serum in 0.1% PBS- and triton X100 for 1h to permeabilise the cells and block non-specific protein-protein interactions. The sections were then incubated with the antibody (ab80255, 1µg/ml) and (ab15640) 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® 568 (red) at a 1/1000 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    See Abreview

References

This product has been referenced in:

  • Lui JC  et al. Differential aging of growth plate cartilage underlies differences in bone length and thus helps determine skeletal proportions. PLoS Biol 16:e2005263 (2018). Read more (PubMed: 30036371) »
  • Loscertales M  et al. Type IV collagen drives alveolar epithelial-endothelial association and the morphogenetic movements of septation. BMC Biol 14:59 (2016). IHC . Read more (PubMed: 27412481) »
See all 2 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Application
Western blot
Sample
Mouse Cell lysate - whole cell (chondrocytes)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
chondrocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Jun 12 2018

Application
Western blot
Sample
Mouse Cell lysate - whole cell (embryonic stem cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
embryonic stem cells
Blocking step
Odyssey blocking buffer, LiCor as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Nov 01 2016

Answer

Thank you for contacting us.

We currently have two rabbit polyclonal antibodies that will detect SMAD 1/5:

Rabbit polyclonal to SMAD1/5/9 - ab80255
Rabbit polyclonal to SMAD 1/5/8/9 - ab13723

We have a number of antibodies that will detect the S463 or S465 phosphorylation on SMAD 1 or SMAD 5:

Rabbit polyclonal to Smad1 (phospho S465)- ab63439
Rabbit monoclonal [EP728(2)AY] to SMAD5 (phospho S463 + S465)- ab76296
Rabbit monoclonal [MMC-1-104-3] to SMAD5 (phospho S463 + S465) - ab92698
Rabbit polyclonal to Smad1 (phospho S465)- ab97689
Rabbit polyclonal to Smad1 (phospho S465)- ab97716

While we only currently carry antibodies raised in rabbit for these targets, you could use our EasyLink conjugation kits to conjugate these antibodies to Cy5.5 or Cy7. These two fluorophores should be compatible with the Odyssey system.

https://www.abcam.com/EasyLink

Read More

Question
Answer

Thank you for contacting us. Of the three antibodies we discussed yesterday, ab80255 is the best choice, given that we know the immunogen is derived from the N terminus, and your detection antibody is directed against the C terminus. The epitope that ab75273 binds to is unmapped, and so I am concerned that it may not be compatible in a sandwich ELISA with your detection antibody if, by chance, it binds to the C terminus. Antibody ab72504 was actually a duplicate entry of ab80255 and has been removed from the catalogue. Ab80255 has not, however, been tested in ELISA to capture native SMAD1. As I expected, it was validated for ELISA against the peptide that was used as the immunogen. It has been validated for IHC, though, so we expect it will be capable of binding the N terminus when SMAD1 is in its native conformation. Another antibody to consider is ab63356. The situation is the same as for ab80255: untested in ELISA against native, endogenous SMAD1, only against the peptide, but it gives a signal in IHC and ICC. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.  

Read More
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Rat Cell (Rat cortex (day P7))
Specification
Rat cortex (day P7)
Fixative
Paraformaldehyde
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted Dec 21 2009

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