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Phospho-Smad2-S465/S467 + Smad3-S423/S425 Rabbit mAb (AP1343)

Tested applications:WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP InhibitionReactivity:Human, Mouse, Rat

Western blot analysis of various lysates, using Phospho-Smad2-S465/S467 + Smad3-S423/S425 Rabbit mAb (AP1343) at1:2000 dilution. HeLa and NIH/3T3 cells were treated by TGF-β3（H） (200 ng/mL) at 37℃ for 30 minutes after serum-starvation overnight.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 60s.

Western blot analysis of various lysates using Phospho-Smad2-S465/S467 + Smad3-S423/S425 Rabbit mAb (AP1343) at 1:1000 dilution. C6 cells were treated by TGF-β3 (200 ng/mL) at 37℃ for 30 minutes after serum-starvation overnight.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 30s.

Overview

Product name	Phospho-Smad2-S465/S467 + Smad3-S423/S425 Rabbit mAb
Catalog No.	AP1343
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG
CloneNo.	ARC56314

Background

The protein encoded by this gene belongs to the SMAD, a family of proteins similar to the gene products of the Drosophila gene 'mothers against decapentaplegic' (Mad) and the C. elegans gene Sma. SMAD proteins are signal transducers and transcriptional modulators that mediate multiple signaling pathways. This protein mediates the signal of the transforming growth factor (TGF)-beta, and thus regulates multiple cellular processes, such as cell proliferation, apoptosis, and differentiation. This protein is recruited to the TGF-beta receptors through its interaction with the SMAD anchor for receptor activation (SARA) protein. In response to TGF-beta signal, this protein is phosphorylated by the TGF-beta receptors. The phosphorylation induces the dissociation of this protein with SARA and the association with the family member SMAD4. The association with SMAD4 is important for the translocation of this protein into the nucleus, where it binds to target promoters and forms a transcription repressor complex with other cofactors. This protein can also be phosphorylated by activin type 1 receptor kinase, and mediates the signal from the activin. Alternatively spliced transcript variants have been observed for this gene. [provided by RefSeq, May 2012]

Immunogen information

Immunogen	A synthetic phosphorylated peptide around S465 & S467 of human SMAD2 (NP_005892.1).
Sequence	SSMS
Gene ID
Swiss Prot
Synonyms	HSPC193; HsT17436; JV15-2; LDS1C; LDS3; MADH3; Phospho-Smad2-S465/S467 + Smad3-S423/S425
Calculated MW	48kDa/52kDa/25kDa/35kDa/43kDa
Observed MW	52kDa/60kDa

Applications

Reactivity	Human, Mouse, Rat
Tested applications	WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP Inhibition
Recommended dilution	WB 1:1000 - 1:5000 ELISA Recommended starting concentration is 1 μg/mL. Please optimize the concentration based on your specific assay requirements.
Storage buffer	Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.05% proclin300, 0.05% BSA, 50% glycerol, pH7.3.
Key application	Western blotting
Positive samples	HeLa treated by TGF-β3, NIH/3T3 treated by TGF-β3, C6 treated by TGF-β3
Cellular location	Cytoplasm, Nucleus.
Customer validation	WB(Rattus norvegicus, Mus musculus, Canis familiaris, Homo sapiens) IHC(Rattus norvegicus)