Immunofluorescence analysis of SMAD7 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 labeled with SMAD7 Rabbit Polyclonal Antibody (420400) at 1ug/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 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® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing Nuclear localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Rat, Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide derived from the C-terminal region of the human|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/million cells|
|Immunocytochemistry (ICC)||1 µg/ml|
|Immunofluorescence (IF)||1 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:50|
|Western Blot (WB)||1-2 µg/ml|
* 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 3 publications below|
Smad7 is a member of the MAD-related family of molecules. MAD-related proteins are a recently identified family of intracellular proteins that are thought to be essential components in the signaling pathways of the serine/threonine kinase receptors of the transforming growth factor beta super family.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Cucurbitacin I Attenuates Cardiomyocyte Hypertrophy via Inhibition of Connective Tissue Growth Factor (CCN2) and TGF- ß/Smads Signalings.
42-0400 was used in western blot to determine the functional role of cucurbitacin I using an in vitro model of cardiac hypertrophy.
|Jeong MH,Kim SJ,Kang H,Park KW,Park WJ,Yang SY,Yang DK||PloS one (10:null)||2015|
Neuropilin-2 Is upregulated in lung cancer cells during TGF-ß1-induced epithelial-mesenchymal transition.
42-0400 was used in western blot to study TGF-beta-driven lung cancer cell epithelial-to-mesenchymal transition and the role played by neuropilin-2.
|Nasarre P,Gemmill RM,Potiron VA,Roche J,Lu X,Barón AE,Korch C,Garrett-Mayer E,Lagana A,Howe PH,Drabkin HA||Cancer research (73:7111)||2013|
|Mouse||Not Cited||Kruppel-like factor KLF10 targets transforming growth factor-beta1 to regulate CD4(+)CD25(-) T cells and T regulatory cells.||Cao Z,Wara AK,Icli B,Sun X,Packard RR,Esen F,Stapleton CJ,Subramaniam M,Kretschmer K,Apostolou I,von Boehmer H,Hansson GK,Spelsberg TC,Libby P,Feinberg MW||The Journal of biological chemistry (284:24914)||2009|