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AB300164

Anti-SMAD1/5/9 antibody [EPR25803-154]

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Rabbit Recombinant Monoclonal SMAD5 antibody. Suitable for WB and reacts with Human, Mouse, Rat samples.

View Alternative Names

MADH5, SMAD5, Mothers against decapentaplegic homolog 5, MAD homolog 5, Mothers against DPP homolog 5, JV5-1, SMAD family member 5, SMAD 5, Smad5, hSmad5

3 Images
Western blot - Anti-SMAD1/5/9 antibody [EPR25803-154] (AB300164)
  • WB

Supplier Data

Western blot - Anti-SMAD1/5/9 antibody [EPR25803-154] (AB300164)

Blocking and diluting buffer and concentration : 5% NFDM/TBST This antibody reacts with human SMAD1, SMAD5, and SMAD9, but does not cross-react with human SMAD2, SMAD3, SMAD6, and SMAD7.

All lanes:

Western blot - Anti-SMAD1/5/9 antibody [EPR25803-154] (ab300164) at 1/1000 dilution

Lane 1:

HEK-293T cells transfected with a human SMAD1 expression vector containi a his-tag, whole cell lysate 10 µg

Lane 2:

HEK-293T cells transfected with a human SMAD2 expression vector containi a his-tag, whole cell lysate, 20 µg

Lane 3:

HEK-293T cells transfected with a human SMAD3 expression vector containi a his-tag, whole cell lysate, 20 µg

Lane 4:

HEK-293T cells transfected with a human SMAD5 expression vector containi a his-tag, whole cell lysate, 10 µg

Lane 5:

His-tagged human SMAD6 recombinant protein 10 ng

Lane 6:

HEK-293T cells transfected with a human SMAD7 expression vector containi a his-tag, whole cell lysate, 20 µg

Lane 7:

HEK-293T cells transfected with a human SMAD9 expression vector containi a his-tag, whole cell lysate, 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

Observed band size: 20 kDa,62 kDa

false

Exposure time: 7.75s

Western blot - Anti-SMAD1/5/9 antibody [EPR25803-154] (AB300164)
  • WB

Supplier Data

Western blot - Anti-SMAD1/5/9 antibody [EPR25803-154] (AB300164)

Blocking and diluting buffer and concentration : 5% NFDM/TBST The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 21220940). This blot was developed using a high sensitivity ECL substrate. The identity of the band at approximately 37 kDa is unknown.

All lanes:

Western blot - Anti-SMAD1/5/9 antibody [EPR25803-154] (ab300164) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 20 µg

Lane 2:

293T (human embryonic kidney epithelial cell) whole cell lysate 20 µg

Lane 3:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 20 µg

Lane 4:

PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate 20 µg

Lane 5:

THP-1 (human monocytic leukemia monocyte) whole cell lysate 20 µg

Lane 6:

Jurkat (human T cell leukemia T lymphocyte) whole cell lysate 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

Observed band size: 52 kDa,54 kDa

false

Exposure time: 147s

Western blot - Anti-SMAD1/5/9 antibody [EPR25803-154] (AB300164)
  • WB

Supplier Data

Western blot - Anti-SMAD1/5/9 antibody [EPR25803-154] (AB300164)

Blocking and diluting buffer and concentration : 5% NFDM/TBST The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 21220940). This blot was developed using a high sensitivity ECL substrate. The identity of the band at approximately 37 kDa is unknown.

All lanes:

Western blot - Anti-SMAD1/5/9 antibody [EPR25803-154] (ab300164) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) transfected with scrambled siRNA control, whole cell lysate 20 µg

Lane 2:

HeLa transfected with siRNA specifically targeting SMAD9, whole cell lysate 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 1/20000 dilution

Observed band size: 52 kDa

false

Exposure time: 3min

  • Carrier free

    Anti-SMAD1/5/9 antibody [EPR25803-154] (BSA and Azide free)

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR25803-154

Isotype

IgG

Carrier free

No

Reacts with

Human, Mouse, Rat

Applications

WB

applications

Immunogen

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

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p> Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "<p></p>" }, "Mouse": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p> Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "<p></p>" }, "Rat": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p> Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "" } } }
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Product details

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.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Preservative: 0.01% Sodium azide Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
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
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Supplementary information

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

SMAD1/5/9 proteins also known as SMAD family members 1 5 and 9 play key mechanical roles in the TGF-beta signaling pathway. These proteins have molecular masses ranging between 52-60 kDa. They are widely expressed in various tissues with notable expression in embryonic tissues and the central nervous system. These proteins operate as intracellular signal transducers and transcriptional modulators blending both cytoplasmic and nuclear functions.
Biological function summary

SMAD1/5/9 proteins serve critical functions in the cellular response to signaling molecules like bone morphogenetic proteins (BMPs). They form part of a complex with receptor-regulated SMADs (R-SMADs) and common-mediator SMAD4 which translocates to the nucleus to regulate the transcription of target genes. These proteins have roles in processes like bone and cartilage development cell growth and apoptosis engaging as vital mediators in various cellular contexts.

Pathways

SMAD1/5/9 proteins are integral components of the BMP signaling pathway which governs numerous biological functions such as differentiation and embryogenesis. These proteins work alongside partners such as SMAD4 to mediate the transcription of genes involved in these intricate processes. Additionally they interact with proteins in the SMAD pathway modulated by TGF-beta and engage in crosstalk with other signaling pathways to coordinate cellular outcomes.

SMAD1/5/9 proteins have connections to pulmonary hypertension and skeletal malformations. Aberrant activity or dysregulation of these proteins relates to the progression of these conditions. In pulmonary hypertension the abnormal regulation of the BMP pathway often through interactions with endothelial cells involves SMAD1/5/9. In skeletal disorders such as congenital skeletal malformations these proteins influence bone formation and maintenance often working in conjunction with other proteins such as SMAD4 whose disruptions can exacerbate disease states.
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Product protocols

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

Transcriptional regulator that plays a role in various cellular processes including embryonic development, cell differentiation, angiogenesis and tissue homeostasis (PubMed : 12064918, PubMed : 16516194). 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 : 9442019). In turn, the hetero-trimeric complex recognizes cis-regulatory elements containing Smad Binding Elements (SBEs) to modulate the outcome of the signaling network (PubMed : 33510867). Non-phosphorylated SMAD5 has a cytoplasmic role in energy metabolism regulation by promoting mitochondrial respiration and glycolysis in response to cytoplasmic pH changes (PubMed : 28675158). Mechanistically, interacts with hexokinase 1/HK1 and thereby accelerates glycolysis (PubMed : 28675158).
See full target information SMAD5

Additional targets

SMAD5,SMAD9
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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
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Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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