Anticorps Recombinant de lapin anti-SMAD4

• Phare
• Validé par KD/KO

SMAD4 Recombinant Antibody for WB, IF/ICC, FC (Intra), ELISA, ChIP-qPCR

Hôte / Isotype

Lapin / IgG

Réactivité testée

Humain, souris

Applications

WB, IF/ICC, FC (Intra), ELISA, ChIP-qPCR

Conjugaison

Non conjugué

CloneNo.

240026G9

N° de cat : 83169-5-RR

Synonymes

240026G9, DPC4, hSMAD4, JIP, MAD homolog 4

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Galerie de données de validation

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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 1:20000 incubated at room temperature for 1.5 hours.

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  WB analysis of HEK-293 using 83169-5-RR

  WB result of SMAD4 antibody (83169-5-RR; 1:6000; incubated at room temperature for 1.5 hours) with sh-Control and sh-SMAD4 transfected HEK-293 cells.

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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 AffiniPure 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 AffiniPure Goat Anti-Rabbit IgG(H+L)(red), or 0.25 ug rabbit IgG isotype control (blue). Cells were fixed and permeabilized with True-Nuclear Transcription Factor Buffer Set.

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  ChIP experiment of HaCaT using 83169-5-RR

  Chromatin was prepared from HaCaT cells either non-treated or treated with TGF-β1 (7 ng/ml, 1 h). Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 20 µg of cross-linked chromatin, 5 µg of SMAD4 (83169-5-RR) or 5 ug of Normal Rabbit IgG (98136-1-RR), and 20 µl of Protein A Magarose Beads. The immunoprecipitated DNA was quantified by real-time PCR.

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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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Applications testées

• Résultats positifs en WB: cellules BxPC-3, cellules HEK-293, cellules MCF-7, cellules NIH/3T3
• Résultats positifs en IF/ICC: cellules HepG2,
• Résultats positifs en FC (Intra): cellules HepG2
• Résultats positifs en ChIP-qPCR: TGF-β1 (7 ng/ml, 1 h) HaCaT cells,

Dilution recommandée

• 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
• CHImmunoprécipitation (IP)-QPCR: CHIP-QPCR : 1:10-1:100

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

Informations sur le produit

83169-5-RR cible SMAD4 dans les applications de WB, IF/ICC, FC (Intra), ELISA, ChIP-qPCR et montre une réactivité avec des échantillons Humain, souris

• Réactivité: Humain, souris
• Hôte / Isotype: Lapin / IgG
• Clonalité: Recombinant
• Type: Anticorps
• Immunogène: SMAD4 Protéine recombinante Ag0299
• Nom complet: SMAD family member 4
• Masse moléculaire calculée: 60 kDa
• Poids moléculaire observé: 63-70 kDa
• Numéro d’acquisition GenBank: BC002379
• Symbole du gène: SMAD4
• Identification du gène (NCBI): 4089
• Conjugaison: Non conjugué
• Forme: Liquide
• Méthode de purification: Protein A purfication
• Tampon de stockage: PBS with 0.02% sodium azide and 50% glycerol
• Conditions de stockage: Stocker à -20°C. Stable pendant un an après l'expédition. L'aliquotage n'est pas nécessaire pour le stockage à -20^oC Les 20ul contiennent 0,1% de BSA.

Informations générales

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.

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