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Phospho-Smad3-S423/S425 Rabbit pAb (AP1263)

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 (AP1263) at 1:1000 dilution. Hela cells were treated by TGF-β (10 ng/ml) at 37℃ for 30 minutes.
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: 90s.

Western blot analysis of various lysates using (AP1263) at 1:1000 dilution. C2C12 cells were treated by TGF-β (10 ng/ml) at 37℃ for 30 minutes.
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: 180s.

ABclonal:Immunohistochemistry - Phospho-Smad3-S423/S425 Rabbit pAb (AP1263)

Immunohistochemistry analysis of paraffin-embedded Human colon carcinoma tissue using Phospho-Smad3-S423/S425 Rabbit pAb (AP1263) at a dilution of 1:200 (40x lens). High pressure antigen retrieval was performed with 0.01 M citrate buffer (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Human lung tissue using Phospho-Smad3-S423/S425 Rabbit pAb (AP1263) at a dilution of 1:200 (40x lens). High pressure antigen retrieval was performed with 0.01 M citrate buffer (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Mouse liver tissue using Phospho-Smad3-S423/S425 Rabbit pAb (AP1263) at a dilution of 1:200 (40x lens). High pressure antigen retrieval was performed with 0.01 M citrate buffer (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Rat colon tissue using Phospho-Smad3-S423/S425 Rabbit pAb (AP1263) at a dilution of 1:200 (40x lens). High pressure antigen retrieval was performed with 0.01 M citrate buffer (pH 6.0) prior to IHC staining.

Overview

Product name	Phospho-Smad3-S423/S425 Rabbit pAb
Catalog No.	AP1263
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG

Background

The SMAD family of proteins are a group of intracellular signal transducer proteins similar to the gene products of the Drosophila gene 'mothers against decapentaplegic' (Mad) and the C. elegans gene Sma. The SMAD3 protein functions in the transforming growth factor-beta signaling pathway, and transmits signals from the cell surface to the nucleus, regulating gene activity and cell proliferation. This protein forms a complex with other SMAD proteins and binds DNA, functioning both as a transcription factor and tumor suppressor. Mutations in this gene are associated with aneurysms-osteoarthritis syndrome and Loeys-Dietz Syndrome 3.

Immunogen information

Immunogen	A phospho synthetic peptide corresponding to residues surrounding S423/S425 of Human SMAD3.
Sequence	Email for sequence
Gene ID
Swiss Prot
Synonyms	LDS3; mad3; LDS1C; MADH3; JV15-2; hMAD-3; hSMAD3; HSPC193; HsT17436; Phospho-Smad3-S423/S425
Calculated MW	48kDa
Observed MW	55kDa

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 1:500 - 1:1000 IHC-P 1:50 - 1:200
Storage buffer	Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.05% proclin300, 50% glycerol, pH7.3.
Key application	Western blotting Immunohistochemistry
Positive samples	HeLa treated by TGF-β, C2C12 treated by TGF-β
Cellular location	Cytoplasm, Nucleus.
Customer validation	WB(Mus musculus, Homo sapiens, Rattus norvegicus) PCR(Mus musculus) WB(Mus musculus) qPCR(Mus musculus)