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Rabbit Recombinant Monoclonal SMAD1 phospho S463 + S465 antibody. Suitable for IP, Dot, WB, ICC/IF and reacts with Mouse, Human, Synthetic peptide samples. Cited in 5 publications.

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Images

Dot Blot - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-20] (AB226821), expandable thumbnail
  • Immunoprecipitation - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-20] (AB226821), expandable thumbnail
  • Western blot - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-20] (AB226821), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-20] (AB226821), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPDotWBICC/IF
Human
Expected
Expected
Tested
Expected
Mouse
Tested
Expected
Tested
Tested
Synthetic peptide
Not recommended
Tested
Not recommended
Not recommended

Tested
Tested

Species
Mouse
Dilution info
1/30
Notes

-

Expected
Expected

Species
Human
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Synthetic peptide
Dilution info
-
Notes

-

Tested
Tested

Species
Synthetic peptide
Dilution info
1/1000
Notes

-

Expected
Expected

Species
Mouse, Human
Dilution info
Use at an assay dependent concentration.
Notes

-

Tested
Tested

Species
Mouse
Dilution info
1/1000
Notes

-

Species
Human
Dilution info
1/1000
Notes

-

Not recommended
Not recommended

Species
Synthetic peptide
Dilution info
-
Notes

-

Tested
Tested

Species
Mouse
Dilution info
1/100
Notes

-

Expected
Expected

Species
Human
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Synthetic peptide
Dilution info
-
Notes

-

Associated Products

Select an associated product type

13 products for Alternative Product

Target data

Function

Transcriptional modulator that plays a role in various cellular processes, including embryonic development, cell differentiation, and tissue homeostasis (PubMed:9335504). Upon BMP ligand binding to their receptors at the cell surface, is phosphorylated by activated type I BMP receptors (BMPRIs) and associates with SMAD4 to form a heteromeric complex which translocates into the nucleus acting as transcription factor (PubMed:33667543). In turn, the hetero-trimeric complex recognizes cis-regulatory elements containing Smad Binding Elements (SBEs) to modulate the outcome of the signaling network (PubMed:33667543). SMAD1/OAZ1/PSMB4 complex mediates the degradation of the CREBBP/EP300 repressor SNIP1. Positively regulates BMP4-induced expression of odontogenic development regulator MSX1 following IPO7-mediated nuclear import (By similarity).

Alternative names

BSP1, MADH1, MADR1, SMAD1, Mothers against decapentaplegic homolog 1, MAD homolog 1, Mothers against DPP homolog 1, JV4-1, Mad-related protein 1, SMAD family member 1, Transforming growth factor-beta-signaling protein 1, SMAD 1, Smad1, hSMAD1, BSP-1

Recommended products

Rabbit Recombinant Monoclonal SMAD1 phospho S463 + S465 antibody. Suitable for IP, Dot, WB, ICC/IF and reacts with Mouse, Human, Synthetic peptide samples. Cited in 5 publications.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR20662-20
Purification technique
Affinity purification Protein A
Specificity

Based on sequence homology this antibody also reacts with Smad5 (phospho S463/S465) and Smad9 (phospho S465/S467).

Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Notes

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 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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

Smad1 also known as Mothers against decapentaplegic homolog 1 (MADH1) is a protein encoded by the SMAD1 gene in humans. It has a molecular mass of approximately 53 kDa and functions as a receptor-regulated Smad protein. Smad1 primarily transmits signals from the bone morphogenetic proteins (BMPs) a group of growth factors and cytokines. Expression of Smad1 occurs in various tissues including the lung heart and kidney. It plays an important role in mediating signaling pathways within different cellular environments.

Biological function summary

Smad1 participates in the transmission of BMP signals from the cell surface to the nucleus ensuring transcriptional regulation of target genes. In the presence of BMPs Smad1 forms complexes with Smad4 upon activation. These Smad complexes then translocate to the nucleus where they regulate gene expression. Smad1 influences cellular responses such as proliferation differentiation and apoptosis. In particular it plays a significant role in bone development and osteogenesis.

Pathways

Smad1 plays an important role within the BMP signaling pathway which is important for early development and tissue homeostasis. Within this pathway BMPs trigger the phosphorylation of Smad1 which then associates with Smad4 to propagate downstream signaling. Another associated pathway includes the TGF-beta signaling pathway where Smads like Smad2 and Smad3 show functional similarities and differences with Smad1. The interactions of Smad1 with related proteins like Smad4 highlight its significant contribution to cellular processes regulated by these pathways.

Associated diseases and disorders

Smad1 associates with various pathological conditions. For instance aberrant Smad1 signaling has implications in cancer particularly in processes like tumor progression and metastasis. Disruptions in the BMP pathway involving Smad1 and Smad4 can influence the onset and development of disorders such as pulmonary hypertension where inappropriate cell growth and remodeling occur. Understanding Smad1's role in these contexts provides insights into potential therapeutic avenues for treatment and management of these conditions.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

4 product images

  • Dot Blot - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-20] (ab226821), expandable thumbnail

    Dot Blot - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-20] (ab226821)

    Dot blot analysis of Smad1 (phospho S463 + S465) labeled with ab226821 at 1/1000 dilution.

    Lane 1: Smad1 (phospho S463/S465) peptide;

    Lane 2: Smad1 (phospho S463) peptide;

    Lane 3: Smad1 (phospho S465) peptide;

    Lane 4: Smad1 peptide (not phosphorylated);

    Lane 5: Smad5 (phospho S463/S465) peptide;

    Lane 6: Smad5 (phospho S463) peptide;

    Lane 7: Smad5 (phospho S465) peptide;

    Lane 9: Smad5 peptide (not phosphorylated).

    Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution was used as secondary antibody.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: 3 minutes.

    Based on sequence homology, this antibody cross-reacts with Smad5 (phospho S463/S465) and Smad9 (phospho S465/S467).

  • Immunoprecipitation - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-20] (ab226821), expandable thumbnail

    Immunoprecipitation - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-20] (ab226821)

    Smad 1 (phospho S463 + S465) was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryo fibroblast cell line) grown in serum-free media overnight, then treated with 50 ng/ml BMP2 for 30 minutes, whole cell lysate with ab226821 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab226821 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/1000 dilution.

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

    Lane 2: ab226821 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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab226821 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: 10 seconds.

    All lanes: Immunoprecipitation - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-20] (ab226821)

    Predicted band size: 52 kDa

  • Western blot - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-20] (ab226821), expandable thumbnail

    Western blot - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-20] (ab226821)

    Exposure time:
    Lanes 1 and 2: 3 minutes.
    Lanes 3 and 4: 30 seconds.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-20] (ab226821) at 1/1000 dilution

    Lane 1: HeLa (human epithelial cell line from cervix adenocarcinoma) grown in serum-free media overnight, whole cell lysate at 20 µg

    Lane 2: HeLa grown in serum-free media overnight, then treated with 100 ng/ml Calyculin A (Calyculin A, protein phosphatase inhibitor ab141784) for 15 minutes, followed by Calyculin A removal and treatment with 100 ng/ml BMP2 for 30 minutes, whole cell lysate at 20 µg

    Lane 3: NIH/3T3 (mouse embryo fibroblast cell line) grown in serum-free media overnight, whole cell lysate at 20 µg

    Lane 4: NIH/3T3 cultured in serum-free media overnight, then treated with 50 ng/ml BMP2 for 30 minutes, whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (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

  • Immunocytochemistry/ Immunofluorescence - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-20] (ab226821), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-20] (ab226821)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryo fibroblast cell line) cells labeling Smad1 (phospho S463 + S465) with ab226821 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Nuclear staining in hBMP2-treated NIH/3T3 cells. Cells were FBS-deprived overnight before treatment with 50 ng/ml hBMP2 for 30 minutes.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.

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