Immunofluorescence analysis of SMAD3 [pSpS423/425] was done on 70% confluent log phase HeLa cells treated with TGF-beta (20ng/mL for 15 min) cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with SMAD3 [pSpS423/425] Rabbit polyclonal Antibody (44246G) at 2µg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor 488 Goat Anti-Rabbit IgG Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (A12381). Panel d is a merged image showing nuclear translocation of SMAD3 [pSpS423/425]. Panel e cytoplasmic localization. Panel f shows no primary antibody control. The images were captured at 20X magnification.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Rat, Non-human primate, Hamster, Mouse, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from a region of human Smad3 that contains serine 423 and serine 425. This sequence is conserved in mouse.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 1mg/ml BSA, 50% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:100-1:500|
|Western Blot (WB)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Genetic deletion of cell division autoantigen 1 retards diabetes-associated renal injury.
44-246G was used in immunohistochemistry to study the role of cell division autoantigen 1 in diabetes-associated renal injury
|Chai Z,Dai A,Tu Y,Li J,Wu T,Wang Y,Hale LJ,Koentgen F,Thomas MC,Cooper ME||Journal of the American Society of Nephrology : JASN (24:1782)||2013|
||Localization of phosphorylated SMAD proteins in granulosa cells, oocytes and oviduct of female mice.||Tian X,Halfhill AN,Diaz FJ||Gene expression patterns : GEP (10:105)||2010|
|Mouse||1:400||Localization of phosphorylated SMAD proteins in granulosa cells, oocytes and oviduct of female mice.||Tian X,Halfhill AN,Diaz FJ||Gene expression patterns : GEP (10:105)||2010|
|Mouse||1:1000||Impaired angiogenic response in the corneas of mice lacking osteopontin.||Fujita N,Fujita S,Okada Y,Fujita K,Kitano A,Yamanaka O,Miyamoto T,Kon S,Uede T,Rittling SR,Denhardt DT,Saika S||Investigative ophthalmology and visual science (51:790)||2010|
|BRCA1 interacts with Smad3 and regulates Smad3-mediated TGF-beta signaling during oxidative stress responses.||Li H,Sekine M,Seng S,Avraham S,Avraham HK||PloS one (4:null)||2009|
|Activin inhibits the human Pit-1 gene promoter through the p38 kinase pathway in a Smad-independent manner.||de Guise C,Lacerte A,Rafiei S,Reynaud R,Roy M,Brue T,Lebrun JJ||Endocrinology (147:4351)||2006|