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Phospho-Smad2 (Ser467) Rabbit mAb

Catalog No.: F3740

Application: Reactivity: Mouse, Rat, Human

Usage Information

Dilution
WB1:1000 IP1:30 IHC1:2000 IF1:50 FCM1:5000

Application
WB, IP, IHC, IF, FCM

Reactivity
Mouse, Rat, Human

Source
Rabbit

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW Observed MW
52 kDa 60 kDa
^*Why do the predicted and actual molecular weights differ? The following reasons may explain differences between the predicted and actual protein molecular weight.

Datasheet & SDS

Biological Description

Specificity
Phospho-Smad2 (Ser467) Rabbit mAb detects endogenous levels of Smad2 only when phosphorylated at Ser 467.

Clone
P3A22

Synonym(s)
MADH2, MADR2, SMAD2, Mothers against decapentaplegic homolog 2, MAD homolog 2, Mothers against DPP homolog 2, JV18-1, Mad-related protein 2, SMAD family member 2, hMAD-2, SMAD 2, Smad2, hSMAD2

Background
Phospho-Smad2 (Ser467) represents the activated, phosphorylated form of Smad2, a receptor-regulated Smad (R-Smad) involved in the Transforming Growth Factor-beta (TGF-β) signaling pathway. Smad2 comprises two conserved domains: the N-terminal Mad homology 1 (MH1) domain, which mediates DNA binding and nuclear import, and the C-terminal Mad homology 2 (MH2) domain, which is essential for receptor interaction and complex formation with other Smad proteins. The Ser467 phosphorylation site, located within the C-terminal SSXS motif of the MH2 domain, is specifically phosphorylated by activated type I TGF-β receptors. This phosphorylation triggers a conformational shift in Smad2 from a closed, autoinhibited state to an open conformation, promoting its release from the receptor and trimerization with Smad4. The resulting Smad2/Smad4 complex translocates into the nucleus, where it cooperates with transcription factors to regulate genes involved in cell proliferation, differentiation, apoptosis, and epithelial-to-mesenchymal transition (EMT). Phosphorylation at Ser467 is thus critical for Smad2 activation and effective TGF-β signal transduction. Loss or mutation of this phosphorylation site disrupts Smad2-Smad4 complex formation, impairing downstream signaling and affecting biological processes such as embryonic development and tumor suppression.

Background

Phospho-Smad2 (Ser467) represents the activated, phosphorylated form of Smad2, a receptor-regulated Smad (R-Smad) involved in the Transforming Growth Factor-beta (TGF-β) signaling pathway. Smad2 comprises two conserved domains: the N-terminal Mad homology 1 (MH1) domain, which mediates DNA binding and nuclear import, and the C-terminal Mad homology 2 (MH2) domain, which is essential for receptor interaction and complex formation with other Smad proteins. The Ser467 phosphorylation site, located within the C-terminal SSXS motif of the MH2 domain, is specifically phosphorylated by activated type I TGF-β receptors. This phosphorylation triggers a conformational shift in Smad2 from a closed, autoinhibited state to an open conformation, promoting its release from the receptor and trimerization with Smad4. The resulting Smad2/Smad4 complex translocates into the nucleus, where it cooperates with transcription factors to regulate genes involved in cell proliferation, differentiation, apoptosis, and epithelial-to-mesenchymal transition (EMT). Phosphorylation at Ser467 is thus critical for Smad2 activation and effective TGF-β signal transduction. Loss or mutation of this phosphorylation site disrupts Smad2-Smad4 complex formation, impairing downstream signaling and affecting biological processes such as embryonic development and tumor suppression.

References
https://pubmed.ncbi.nlm.nih.gov/25935112/ https://pubmed.ncbi.nlm.nih.gov/11223879/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

Tel: +1-832-582-8158 Ext:3
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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%