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PDP - Template Name: Monoclonal Antibody
PDP - Template ID: *******c5e4b77
R Recombinant
Recombinant: Superior lot-to-lot consistency, continuous supply, and animal-free manufacturing.

SMAD3 (C67H9) Rabbit mAb #9523

Filter:
  • WB
  • IP
  • IF
  • F
  • ChIP
Western Blotting Image 1: SMAD3 (C67H9) Rabbit mAb
Western blot analysis of extracts from control HeLa cells (lane 1) or HeLa cells with an apparent in-frame truncation mutation in the gene encoding SMAD3 (lane 2) using SMAD3 (C67H9) Rabbit mAb #9523 (upper) or β-actin (D6A8) Rabbit mAb #8457 (lower). The change in SMAD3 molecular weight in the mutated HeLa cells is consistent with an in-frame deletion.

To Purchase # 9523

Cat. # Size Qty. Price
9523T 20 µl
$145
9523S 100 µl
$339

Supporting Data

REACTIVITY H M R Mk
SENSITIVITY Endogenous
MW (kDa) 52
Source/Isotype Rabbit IgG
Application Key:
  • WB-Western Blotting 
  • IP-Immunoprecipitation 
  • IF-Immunofluorescence 
  • F-Flow Cytometry 
  • ChIP-Chromatin Immunoprecipitation 
Species Cross-Reactivity Key:
  • H-Human 
  • M-Mouse 
  • R-Rat 
  • Mk-Monkey 
  • Related Products
  • Conjugates

Product Information

Product Usage Information

For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunofluorescence (Immunocytochemistry) 1:100 - 1:200
Flow Cytometry (Fixed/Permeabilized) 1:100
Chromatin IP 1:50
Chromatin IP-seq 1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #44223.

Protocol

Specificity / Sensitivity

SMAD3 (C67H9) Rabbit mAb detects endogenous levels of total SMAD3 protein. No cross reactivity was detected with other family members.

Species Reactivity:

Human, Mouse, Rat, Monkey

The antigen sequence used to produce this antibody shares 100% sequence homology with the species listed here, but reactivity has not been tested or confirmed to work by CST. Use of this product with these species is not covered under our Product Performance Guarantee.

Species predicted to react based on 100% sequence homology:

Xenopus, Zebrafish, Bovine

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues at the amino terminus of SMAD3.

Background

Members of the SMAD family of signal transduction molecules are components of a critical intracellular pathway that transmit TGF-β signals from the cell surface into the nucleus. Three distinct classes of SMADs have been defined: the receptor-regulated SMADs (R-SMADs), which include SMAD1, 2, 3, 5, and 9; the common-mediator SMAD (co-SMAD), SMAD4; and the antagonistic or inhibitory SMADs (I-SMADs), SMAD6 and 7 (1-5). Activated type I receptors associate with specific R-SMADs and phosphorylate them on a conserved carboxy-terminal SSXS motif. The phosphorylated R-SMADs dissociate from the receptor and form a heteromeric complex with SMAD4, initiating translocation of the heteromeric SMAD complex to the nucleus. Once in the nucleus, SMADs recruit a variety of DNA binding proteins that function to regulate transcriptional activity (6-8).

Following stimulation by TGF-β, Smad2 and Smad3 become phosphorylated at their carboxyl termini (Ser465 and 467 on Smad2; Ser423 and 425 on Smad3) by TGF-β Receptor I. Phosphorylated Smad 2/3 can complex with Smad4, translocate to the nucleus and regulate gene expression (9-11).

Pathways

Explore pathways related to this product.


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