AB236156

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

RabMAb
Recombinant
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(1 Publication)

Rabbit Recombinant Monoclonal SMAD1 phospho S463 + S465 antibody. Carrier free. Suitable for IP, Dot, WB and reacts with Mouse, Human, Synthetic peptide samples. Cited in 1 publication.

View 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

3 Images

Lab

Western blot - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] - BSA and Azide free (AB236156)

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).

All lanes:

Western blot - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] (<a href='/en-us/products/primary-antibodies/smad1-phospho-s463-s465-antibody-epr20662-29-ab214423'>ab214423</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

HeLa grown in serum-free media overnight, then treated with 100ng/ml Calyculin A (<a href='/en-us/products/biochemicals/calyculin-a-protein-phosphatase-inhibitor-ab141784'>ab141784</a>) for 15 minutes, Calyculin A was removed, followed by treatment with 100ng/ml BMP2 for 30 minutes, whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

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

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 52 kDa

Observed band size: 60 kDa

true

Exposure time: 10s

Supplier Data

Immunoprecipitation - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] - BSA and Azide free (AB236156)

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).

All lanes:

Immunoprecipitation - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] (<a href='/en-us/products/primary-antibodies/smad1-phospho-s463-s465-antibody-epr20662-29-ab214423'>ab214423</a>)

Predicted band size: 52 kDa

Observed band size: 60 kDa

true

Exposure time: 30s

Supplier Data

Dot Blot - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] - BSA and Azide free (AB236156)

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).

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR20662-29

Isotype

IgG

Carrier free

Yes

Reacts with

Mouse, Human

Applications

Dot, WB, IP

applications

Immunogen

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

Specificity

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

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "Dot" : {"fullname" : "Dot Blot", "shortname":"Dot"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "IP-species-checked": "guaranteed", "IP-species-dilution-info": "", "IP-species-notes": "", "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "" }, "Mouse": { "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "", "IP-species-notes": "", "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "guaranteed", "WB-species-dilution-info": "", "WB-species-notes": "" }, "Synthetic peptide": { "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "Dot-species-checked": "testedAndGuaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "" } } }

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Product details

ab236156 is the carrier-free version of ab214423.

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

What are the advantages of a recombinant monoclonal antibody?
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.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. 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.

Compatibility
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.

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Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

pH: 7.2 - 7.4 Constituents: PBS

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

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Supplementary information

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

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.

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.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

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Target data

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).

See full target information SMAD1 phospho S463 + S465

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Publications (1)

Recent publications for all applications. Explore the full list and refine your search

Nature communications 13:7099 PubMed36402816

2022

The non-catalytic role of DNA polymerase epsilon in replication initiation in human cells.

Applications

Unspecified application

Species

Unspecified reactive species

Sameera Vipat,Dipika Gupta,Sagun Jonchhe,Hele Anderspuk,Eli Rothenberg,Tatiana N Moiseeva

View all publications

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

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

Western blot - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] - BSA and Azide free (AB236156)

Immunoprecipitation - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] - BSA and Azide free (AB236156)

Dot Blot - Anti-Smad1 (phospho S463 + S465) antibody [EPR20662-29] - BSA and Azide free (AB236156)

Key facts

Host species

Clonality

Clone number

Isotype

Carrier free

Reacts with

Applications

Immunogen

Specificity

Reactivity data

Product details

Properties and storage information

Form

Purification technique

Storage buffer

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Publications (1)

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