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Immunofluorescence analysis of Phospho-SMAD2 pSer465 / pSer467 Antibody was done on 70% confluent log phase HeLa 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 treated with TGF-beta1(20ng/mL) for 15 minutes and labelled with Phospho-SMAD2 pSer465 / pSer467 Antibody(44244G) at 1µg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa flour 488 Goat Anti-Rabbit IgG Secondary Antibody (A11008) at a dilution of 1:400 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (A12381). Panel d is a merged image showing Nuclear localization. Panel e showing merged image of untreated cells with less nuclear signals Panel f is a no primary antibody control. The images were captured at 40X magnification.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Mouse, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from a region of human Smad2 that contains serines 465 and 467. The sequence is conserved in mouse and rat.|
|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 *|
|ChIP assay (ChIP)||10µl|
|Flow Cytometry (Flow)||3-5 µg/million cells|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20-1:200|
|Western Blot (WB)||1:500-1:2000|
* 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.
|Western Blot (WB)||See 2 publications below|
SMAD2, also known as MADH2 or MAD2 regulates multiple cellular processes, such as cell proliferation, apoptosis, and differentiation. Smad2 interacts with the TGF-beta receptors through its interaction with the SMAD anchor for receptor activation into the nucleus is a central event in TGF beta signaling. Phosphorylation of threonine 8 in the calmodulin-binding region of the MH1 domain by extracellular signalregulated kinase 1 (ERK1) enhances Smad2 transcriptional activity, which is negatively regulated by calmodulin.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
BMP signaling controls muscle mass.
44-244G was used in western blot to demonstrate that bone morphogenetic protein signaling, acting through Smad, Smad5 and Smad8, is the fundamental hypertrophic signal in mice.
|Sartori R,Schirwis E,Blaauw B,Bortolanza S,Zhao J,Enzo E,Stantzou A,Mouisel E,Toniolo L,Ferry A,Stricker S,Goldberg AL,Dupont S,Piccolo S,Amthor H,Sandri M||Nature genetics (45:1309)||2013|
The significance of a Cripto-1 positive subpopulation of human melanoma cells exhibiting stem cell-like characteristics.
44-244G was used in western blot to investigate the role of cell surface CR- expression in human melanoma cells.
|Strizzi L,Margaryan NV,Gilgur A,Hardy KM,Normanno N,Salomon DS,Hendrix MJ||Cell cycle (Georgetown, Tex.) (12:1450)||2013|
hMAD-2; hSMAD2; MAD homolog 2; mad-related protein 2; MADH2; MADR2; mother against DPP homolog 2; mothers against decapentaplegic homolog 2; Mothers against DPP homolog 2; pSMAD2; Sma- and Mad-related protein 2; SMAD 2; SMAD, mothers against DPP homolog 2; SMAD2
7120426M23Rik; hMAD-2; hSMAD2; JV18; JV18-1; MADH2; MADR2; mMad2; Smad-2; SMAD2