Anti-ATP1A3 antibody [XVIF9-G10] (ab2826)

Samples were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel. Resolved proteins were then transferred onto a Nitrocellulose membrane by iBlot® 2 Dry Blotting System. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit.

Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human prostate carcinoma tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at 1/50 dilution with ab2826 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human colon tissue tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at 1/200 dilution with ab2826 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human tonsil tissue tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at 1/200 dilution with ab2826 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Immunofluorescent analysis of Sodium/Potassium ATPase alpha-3 using ab2826 shows staining in C6 glioma cells. Sodium/Potassium ATPase alpha-3 staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing Sodium/Potassium ATPase alpha-3 ab2826 at a dilution of 1:20 over night at 4 °C washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

Immunofluorescent analysis of Sodium/Potassium ATPase alpha-3 using ab2826 shows staining in U251 glioma cells. Sodium/Potassium ATPase alpha-3 staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing Sodium/Potassium ATPase alpha-3 ab2826 at a dilution of 1:20 over night at 4 °C washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

Immunofluorescent analysis of Sodium/Potassium ATPase alpha-3 using ab2826 shows staining in HeLa cells. Sodium/Potassium ATPase alpha-3 staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing Sodium/Potassium ATPase alpha-3 ab2826 at a dilution of 1:20 over night at 4°C washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.