SMAD4 Polyclonal antibody

• Featured Product
• KD/KO Validated

SMAD4 Polyclonal Antibody for WB, IHC, IP, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat and More (4)

Applications

WB, IHC, IF, IP, CoIP, ChIP, ELISA

Conjugate

Unconjugated

Publications

91

Cat no : 10231-1-AP

Synonyms

Mothers against DPP homolog 4, MADH4, MAD homolog 4, JIP, hSMAD4

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Validation Data Gallery

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  WB analysis using 10231-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 10231-1-AP (SMAD4 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  WB analysis of HEK-293 using 10231-1-AP

  WB result of SMAD4 antibody (10231-1-AP; 1:1000; incubated at room temperature for 1.5 hours) with sh-Control and sh-SMAD4 transfected HEK-293 cells.

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  WB analysis of NIH/3T3 using 10231-1-AP

  NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 10231-1-AP (SMAD4 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of mouse liver using 10231-1-AP

  mouse liver tissue were subjected to SDS PAGE followed by western blot with 10231-1-AP (SMAD4 antibody) at dilution of 1:600 incubated at room temperature for 1.5 hours.

• 

  IP experiment of HeLa using 10231-1-AP

  IP result of anti-SMAD4 (IP:10231-1-AP, 4ug; Detection:10231-1-AP 1:1500) with HeLa cells lysate 1320 ug.

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  IHC staining of human cervical cancer using 10231-1-AP

  Immunohistochemical analysis of paraffin-embedded human cervical cancer tissue slide using 10231-1-AP (SMAD4 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of rat small intestine using 10231-1-AP

  Immunohistochemical analysis of paraffin-embedded rat small intestine tissue slide using 10231-1-AP (SMAD4 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human skin cancer using 10231-1-AP

  Immunohistochemical analysis of paraffin-embedded human skin cancer tissue slide using 10231-1-AP (SMAD4 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse pancreas using 10231-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue slide using 10231-1-AP (SMAD4 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse pancreas using 10231-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue slide using 10231-1-AP (SMAD4 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of rat small intestine using 10231-1-AP

  Immunohistochemical analysis of paraffin-embedded rat small intestine tissue slide using 10231-1-AP (SMAD4 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human tonsillitis using 10231-1-AP

  Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 10231-1-AP (SMAD4 antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human tonsillitis using 10231-1-AP

  Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 10231-1-AP (SMAD4 antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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Tested Applications

• Positive WB detected in: COLO 320 cells, HEK-293 cells, NIH/3T3 cells, mouse liver tissue, HCT 116 cells, HEK-293T cells, Neuro-2a cells
• Positive IP detected in: HeLa cells
• Positive IHC detected in: human skin cancer tissue, human tonsillitis tissue, human cervical cancer tissue, mouse pancreas tissue, rat small intestine tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0

Recommended dilution

• Western Blot (WB): WB : 1:500-1:2000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:500-1:2000

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• KD/KO: See 5 publications below
• WB: See 78 publications below
• IHC: See 10 publications below
• IF: See 7 publications below
• IP: See 4 publications below
• CoIP: See 1 publications below
• ChIP: See 4 publications below

Product Information

10231-1-AP targets SMAD4 in WB, IHC, IF, IP, CoIP, ChIP, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, rat, pig, bovine, hamster, goat
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: SMAD4 fusion protein Ag0299
• Full Name: SMAD family member 4
• Calculated Molecular Weight: 60 kDa
• Observed Molecular Weight: 63-70 kDa
• GenBank Accession Number: BC002379
• Gene Symbol: SMAD4
• Gene ID (NCBI): 4089
• RRID: AB_2193323
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Mammalian homologs of the Drosophila Mad gene include Smad1, Smad2, Smad3, Smad4 (DPC4), Smad5, Smad6, Smad7 and Smad8. Smad1 and Smad5 are effectors of BMP2 and BMP4 function while Smad2 and Smad3 are involved in TGF and activin-mediated growth modulation. Smad4 has been shown to mediate all of the above activities through interaction with various Smad family members . Smad6 and Smad7 regulate the response to activin/ TGF signaling by interfering with TGF -mediated phosphorylation of other Smad family members. This antibody is a rabbit polyclonal antibody raised against an internal region of human SMAD4.

Publications

Species	Application	Title
human	WB	Nat Commun; KMT2A associates with PHF5A-PHF14-HMG20A-RAI1 subcomplex in pancreatic cancer stem cells and epigenetically regulates their characteristics; Authors - Mai Abdel Mouti
human	WB,IP,IF	Transl Res; Deubiquitinase USP13 promotes extracellular matrix expression by stabilizing Smad4 in lung fibroblast cells.; Authors - Xinxin Liao ; KD Validated
human	WB	Theranostics; PMID: 28839463; Authors - Junlong Zhuang
rat	WB,IF	Acta Pharmacol Sin; Role of thrombospondin-1 in high-salt-induced mesenteric artery endothelial impairment in rats; Authors - Fang-Fang Xu ; KD Validated
human	WB	EMBO Rep; Mouse endothelial OTUD1 promotes angiotensin II-induced vascular remodeling by deubiquitinating SMAD3; Authors - Zhuqi Huang
human	WB	Stem Cell Reports; Gene Correction Reverses Ciliopathy and Photoreceptor Loss in iPSC-Derived Retinal Organoids from Retinitis Pigmentosa Patients.; Authors - Wen-Li Deng