SMAD4 Recombinant antibody

SMAD4 Uni-rAb^TM Recombinant Antibody for WB, IF/ICC, FC (Intra), ELISA

Host / Isotype

Rabbit / IgG

Reactivity

Human, mouse

Applications

WB, IF/ICC, FC (Intra), ELISA

Conjugate

Unconjugated

CloneNo.

240026G9

Cat no : 83169-5-RR

Synonyms

DPC4, hSMAD4, JIP, MAD homolog 4, MADH4, Mothers against DPP homolog 4, SMAD 4, SMAD family member 4, SMAD4

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Validation Data Gallery

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  WB analysis using 83169-5-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 83169-5-RR (SMAD4 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  IF Staining of HepG2 using 83169-5-RR

  Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using SMAD4 antibody (83169-5-RR, Clone: 240026G9 ) at dilution of 1:250 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2).

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  FC experiment of HepG2 using 83169-5-RR

  1x10^6 HepG2 cells were intracellularly stained with 0.25 ug SMAD4 Recombinant antibody (83169-5-RR, Clone:240026G9) and APC-Conjugated Goat Anti-Rabbit IgG(H+L)(red), or 0.25 ug Rabbit IgG control Rabbit PolyAb (30000-0-AP) (blue). Cells were fixed and permeabilized with True-Nuclear Transcription Factor Buffer Set.

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  Affinity and Kinetic Characterization of 83169-5-RR

  Biolayer interferometry (BLl) kinetic assays of 83169-5-RR against Human SMAD4 were performed. The affinity constant is below 1 pM.

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Tested Applications

• Positive WB detected in: HeLa cells, HepG2 cells, NIH/3T3 cells, A549 cells, PC-3 cells, MCF-7 cells
• Positive IF/ICC detected in: HepG2 cells
• Positive FC (Intra) detected in: HepG2 cells
• Positive FC detected in: HepG2 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:125-1:500
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.25 ug per 10^6 cells in a 100 µl suspension
• Flow Cytometry (FC): FC : 0.25 ug per 10^6 cells in a 100 µl suspension

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.

Product Information

The immunogen of 83169-5-RR is SMAD4 Fusion Protein expressed in E. coli.

• Tested Reactivity: Human, mouse
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• 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
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purfication
• 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.