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Phospho-Smad3-T179 Rabbit pAb (AP0554)

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 lysates from HeLa cells, using Phospho-Smad3-T179 Rabbit pAb (AP0554). HeLa cells were treated by TNF-α (20 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% BSA.

Western blot analysis of lysates from PC-12 cells, using Phospho-Smad2-S465/467/-Smad3-S423/425 pAb (AP0554) at 1:1000 dilution. PC-12 cells were treated by TNF-α (20 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% BSA.
Detection: ECL Basic Kit (RM00020).
Exposure time: 30s.

ABclonal:Immunohistochemistry - Phospho-Smad3-T179 Rabbit pAb (AP0554)

Immunohistochemistry analysis of paraffin-embedded Human breast using Phospho-Smad3-T179 Rabbit pAb (AP0554) at dilution of 1:200 (40x lens). Microwave antigen retrieval performed with 0.01M Tris/EDTA Buffer (pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Human colon using Phospho-Smad3-T179 Rabbit pAb (AP0554) at dilution of 1:200 (40x lens). Microwave antigen retrieval performed with 0.01M Tris/EDTA Buffer (pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Mouse kidney using Phospho-Smad3-T179 Rabbit pAb (AP0554) at dilution of 1:200 (40x lens). Microwave antigen retrieval performed with 0.01M Tris/EDTA Buffer (pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Rat liver using Phospho-Smad3-T179 Rabbit pAb (AP0554) at dilution of 1:200 (40x lens). Microwave antigen retrieval performed with 0.01M Tris/EDTA Buffer (pH 9.0) prior to IHC staining.

Overview

Product name	Phospho-Smad3-T179 Rabbit pAb
Catalog No.	AP0554
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 synthetic phosphorylated peptide around T179 of human Smad3 (NP_005893.1).
Sequence	PETPP
Gene ID
Swiss Prot
Synonyms	LDS3; mad3; LDS1C; MADH3; JV15-2; hMAD-3; hSMAD3; HSPC193; HsT17436; Phospho-Smad3-T179
Calculated MW	48kDa
Observed MW	48kDa

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 - 1:2000 IHC-P 1:50 - 1:200 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 0.09% Sodium azide, 50% glycerol, pH7.3.
Key application	Western blotting Immunohistochemistry
Positive samples	HeLa treated by TNF-α, PC-12 treated by TNF-α
Cellular location	Cytoplasm, Nucleus.
Customer validation	WB(Capra hircus, Mus musculus, Homo sapiens) IF(Mus musculus)