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 functions as a transcriptional modulator activated by transforming growth factor-beta and is thought to play a role in the regulation of carcinogenesis. [provided by RefSeq, Apr 2009]
Catalog No
A0031
Reactivity
Human, Mouse, Rat
Applications
IF/ICC, WB, IHC-p, ELISA
Modification
Phospho Specific
Source
Polyclonal Rabbit
Dilution
IF: 1:50-200 Western Blot: 1/500 - 1/2000. Immunohistochemistry: 1/100 - 1/300. ELISA: 1/40000. Not yet tested in other applications.
Purification
The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen.
Concentration
1 mg/ml
Storage and Stability
-20°C/1 year
Other Name
SMAD3; MADH3; Mothers against decapentaplegic homolog 3; MAD homolog 3; Mad3; Mothers against DPP homolog 3; hMAD-3; JV15-2; SMAD family member 3; SMAD 3; Smad3; hSMAD3
Molecular Weight (Da)
48081
Gene Name
SMAD3
Protein Name
Mothers against decapentaplegic homolog 3
Human Gene ID
4088
Human Swiss Prot No.
P84022
Immunogen
The antiserum was produced against synthesized peptide derived from human Smad3 around the phosphorylation site of Ser425. AA range:376-425
Specificity
Phospho-Smad3 (S425) Polyclonal Antibody detects endogenous levels of Smad3 protein only when phosphorylated at S425.
Formulation
Liquid in PBS containing 50% glycerol, 0.5% BSA and 0.02% sodium azide.
Western blot analysis of lysates from 1) L929, 2) RAT MUSLE,3)MOUSE-BRAIN, 4) RAT- KIDNEY cells, (Green) primary antibody was diluted at 1:1000, 4°over night, secondary antibody(cat:SA0439)was diluted at 1:10000, 37° 1hour. (Red) Actin β Monoclonal Antibody(5B7) (cat:N1292) antibody was diluted at 1:5000 as loading control, 4° over night,secondary antibody(cat:SA0436)was diluted at 1:10000, 37° 1hour.
Western blot analysis of lysates from NIH/3T3 cells treated with IGF-I 100ng/ml 10', using Smad3 (Phospho-Ser425) Antibody. The lane on the right is blocked with the phospho peptide.
Immunohistochemistry analysis of paraffin-embedded human breast carcinoma, using Smad3 (Phospho-Ser425) Antibody. The picture on the right is blocked with the phospho peptide.
Enzyme-Linked Immunosorbent Assay (Phospho-ELISA) for Immunogen Phosphopeptide (Phospho-left) and Non-Phosphopeptide (Phospho-right), using Smad3 (Phospho-Ser425) Antibody
Immunohistochemical analysis of paraffin-embedded Mouse-lung tissue. 1,Smad3 (phospho Ser425) Polyclonal Antibody was diluted at 1:200(4°C,overnight). 2, Sodium citrate pH 6.0 was used for antibody retrieval(>98°C,20min). 3,Secondary antibody was diluted at 1:200(room temperature, 30min). Negative control was used by secondary antibody only.
Immunohistochemical analysis of paraffin-embedded Human-uterus tissue. 1,Smad3 (phospho Ser425) Polyclonal Antibody was diluted at 1:200(4°C,overnight). 2, Sodium citrate pH 6.0 was used for antibody retrieval(>98°C,20min). 3,Secondary antibody was diluted at 1:200(room temperature, 30min). Negative control was used by secondary antibody only.
Immunofluorescence analysis of Hela cell. 1,Smad3 (phospho Ser425) Polyclonal Antibody(red) was diluted at 1:200(4° overnight). ATG5 mouse Monoclonal Antibody(3C7)(green) was diluted at 1:200(4° overnight). 2, Goat Anti Rabbit Alexa Fluor 594 Catalog:SA0837 was diluted at 1:1000(room temperature, 50min). Goat Anti Mouse Alexa Fluor 488 Catalog:SA0662 was diluted at 1:1000(room temperature, 50min).