SMAD4 Rekombinanter Antikörper

• Featured Product
• KD/KO Validated

SMAD4 Rekombinant Antikörper für WB, IF/ICC, FC (Intra), ELISA, ChIP-qPCR

Wirt / Isotyp

Kaninchen / IgG

Getestete Reaktivität

human, Maus

Anwendung

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

Konjugation

Unkonjugiert

CloneNo.

240026G9

Kat-Nr. : 83169-5-RR

Synonyme

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

---

• 

  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.

• 

  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.

• 

  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).

• 

  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.

• 

  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.

• 

  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.

---

Geprüfte Anwendungen

• Erfolgreiche Detektion in WB: BxPC-3-Zellen, HEK-293-Zellen, MCF-7-Zellen, NIH/3T3-Zellen
• Erfolgreiche Detektion in IF/ICC: HepG2-Zellen
• Erfolgreiche Detektion in FC (Intra): HepG2-Zellen
• Erfolgreiche Detektion in ChIP-qPCR: TGF-β1 (7 ng/ml, 1 h) HaCaT cells

Empfohlene Verdünnung

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

Produktinformation

83169-5-RR bindet in WB, IF/ICC, FC (Intra), ELISA, ChIP-qPCR SMAD4 und zeigt Reaktivität mit human, Maus

• Getestete Reaktivität: human, Maus
• Wirt / Isotyp: Kaninchen / IgG
• Klonalität: Rekombinant
• Typ: Antikörper
• Immunogen: SMAD4 fusion protein Ag0299
• Vollständiger Name: SMAD family member 4
• Berechnetes Molekulargewicht: 60 kDa
• Beobachtetes Molekulargewicht: 63-70 kDa
• GenBank-Zugangsnummer: BC002379
• Gene symbol: SMAD4
• Gene ID (NCBI): 4089
• Konjugation: Unkonjugiert
• Form: Liquid
• Reinigungsmethode: Protein A purfication
• Lagerungspuffer: PBS with 0.02% sodium azide and 50% glycerol
• Lagerungsbedingungen: Bei -20°C lagern. Nach dem Versand ein Jahr lang stabil Aliquotieren ist bei -20^oC Lagerung nicht notwendig. 20ul Größen enthalten 0,1% BSA.

Hintergrundinformationen

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.