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Immunohistochemistry Validation of SCF in Mouse Brain Tissue Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-SCF antibody (5165) at 2.5 &956,g/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT, antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4 &730,C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin. |
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Western Blot Validation with Recombinant ProteinLoading: 30 ng of human SCF recombinant protein per lane.Antibodies: SCF 5165 (Lane 1: 1 &956,g/mL and Lane 2: 2 &956,g/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.Observed at around 20kD. |
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Immunofluorescence Validation of SCF in Human Brain TissueImmunofluorescent analysis of 4% paraformaldehyde-fixed human brain tissue labeling SCF with 5165 at 20 &956,g/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red). |
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Western Blot Validation in Cell Lines and Tissues of Human, Mouse and RatLoading: 15 &956,g of lysates per lane.Antibodies: SCF 5165 (1 &956,g/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. |
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Regulation Validation of SCF in Streptozotocin (STZ)-induced Diabetic Mice (Zhang et al., 2016) WB analysis showed protein expression level of SCF detected by anti-SCF antibody (5165) in gastric smooth muscle was significantly decreased in STZ-induced diabetic mice as compared to the control. |
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Regulated Expression Validation of SCF in Gastric Smooth Muscle Cells (GSMCs) of Mice (Zhang et al., 2016) WB analysis of protein expression level of SCF detected by anti-SCF antibody (5165). GSMCs were treated with PI3K inhibitor (LY294002), AT1R inhibitor (ZD7155) and AT2R inhibitor (PD123319) before Ang II treatment. Ang II (10-8mol/L) significantly increased SCF protein expression, which was reduced by treatment of LY294002 and ZD7155. |
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Regulated Expression Validation of SCF in GSMCs of Normal Mice (Wu et al., 2013) WB analysis of protein expression level of SCF detected by anti-SCF antibody (5165). GSMCs were treated with C-type natriuretic peptide (CNP) at different doses for 48hr. CNP(10-7mol/L and 10-6mol/L) significantly decreased SCF protein expression level as compared to the control group. |