SMAD4 Recombinant monoclonal antibody, PBS Only (Capture)

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  WB analysis using 83169-5-RR (same clone as 83169-5-PBS)

  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. This data was developed using the same antibody clone with 83169-5-PBS in a different storage buffer formulation.

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  WB analysis of HEK-293 using 83169-5-RR (same clone as 83169-5-PBS)

  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. This data was developed using the same antibody clone with 83169-5-PBS in a different storage buffer formulation.

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  Sandwich ELISA sample test of MP00267-4

  Serum of sixteen individual healthy human donors were measured. The human SMAD4 concentration of detected samples was determined to be 47.3 pg/mL with a range of ND - 148.8 pg/mL.

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  Sandwich ELISA sample test of MP00267-4

  The mean human SMAD4 concentration was determined to be 9,362.0 pg/mL in HepG2 cell extract based on a 6.0 mg/mL extract load and 4,272.2 pg/mL in HL-60 cell extract based on a 3 mg/mL extract load.

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  Sandwich ELISA standard curve of MP00267-4

  Sandwich ELISA standard curve of MP00267-4, Human SMAD4 Monoclonal Matched Antibody Pair - PBS only. 83169-5-PBS was coated to a plate as the capture antibody and incubated with serial dilutions of standard Ag0299. 83169-6-PBS was HRP conjugated as the detection antibody. Range: 39.1-2500 pg/mL.

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  IF Staining of HepG2 using 83169-5-RR (same clone as 83169-5-PBS)

  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). This data was developed using the same antibody clone with 83169-5-PBS in a different storage buffer formulation.

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  FC experiment of HepG2 using 83169-5-RR (same clone as 83169-5-PBS)

  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. This data was developed using the same antibody clone with 83169-5-PBS in a different storage buffer formulation.

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  ChIP experiment of HaCaT using 83169-5-RR (same clone as 83169-5-PBS)

  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. This data was developed using the same antibody clone with 83169-5-PBS in a different storage buffer formulation.

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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.

• Featured Product
• KD/KO Validated

SMAD4 Uni-rAb^® Recombinant Antibody for WB, IF/ICC, FC (Intra), Sandwich ELISA, Indirect ELISA, ChIP-qPCR

Cat No. 83169-5-PBS

Clone No.240026G9

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse

Applications

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

Synonyms

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

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Product Information

83169-5-PBS targets SMAD4 as part of a matched antibody pair:

MP00267-4: 83169-5-PBS capture and 83169-6-PBS detection (validated in Sandwich ELISA)

Unconjugated rabbit recombinant monoclonal antibody in PBS only (BSA and azide free) storage buffer at a concentration of 1 mg/mL, ready for conjugation. Created using Proteintech’s proprietary in-house recombinant technology. Recombinant production enables unrivalled batch-to-batch consistency, easy scale-up, and future security of supply.

This conjugation ready format makes antibodies ideal for use in many applications including: ELISAs, multiplex assays requiring matched pairs, mass cytometry, and multiplex imaging applications.Antibody use should be optimized by the end user for each application and assay.

• Tested Reactivity: human, mouse
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen:

  CatNo: Ag0299

  Product name: Recombinant human SMAD4 protein

  Source: e coli.-derived, PGEX-4T

  Tag: GST

  Domain: 13-227 aa of BC002379

  Sequence: NDACLSIVHSLMCHRQGGESETFAKRAIESLVKKLKEKKDELDSLITAITTNGAHPSKCVTIQRTLDGRLQVAGRKGFPHVIYARLWRWPDLHKNELKHVKYCQYAFDLKCDSVCVNPYHYERVVSPGIDLSGLTLQSNAPSSMMVKDEYVHDFEGQPSLSTEGHSIQTIQHPPSNRASTETYSTPALLAPSESNATSTANFPNIPVASTSQPAS

• 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
• UNIPROT ID: Q13485
• Storage Buffer: PBS only, pH 7.3.
• Storage Conditions: Store at -80°C.

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.