All lanes : Anti-SERCA1 ATPase antibody [VE121G9] (ab2819) at 1/2000 dilution

Lane 1 : Human skeletal muscle.
Lane 2 : Rat skeletal muscle.
Lane 3 : Mouse skeletal muscle

Lysates/proteins at 25 µg per lane.

Secondary
All lanes : HRP-conjugated secondary antibody

Developed using the ECL technique.

Predicted band size: 110 kDa

Immunocytochemistry/Immunofluorescence analysis of SERCA1 ATPase shows staining in A2058 cells. SERCA1 ATPase 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 incubated without (control) or with or ab2819 (1:20) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated goat anti-mouse IgG secondary antibody. Images were taken at 60X magnification.

Immunocytochemistry/Immunofluorescence analysis of SERCA1 ATPase shows staining in HeLa cells. SERCA1 ATPase 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 incubated without (control) or with or ab2819 (1:20) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated goat anti-mouse IgG secondary antibody. Images were taken at 60X magnification.

Immunocytochemistry/Immunofluorescence analysis of SERCA1 ATPase shows staining in U251 cells. SERCA1 ATPase 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 incubated without (control) or with or ab2819 (1:20) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated goat anti-mouse IgG secondary antibody. Images were taken at 60X magnification.

Anti-SERCA1 ATPase antibody [VE121G9] (ab2819) at 1/500 dilution + Skeletal Muscle (Mouse) Tissue Lysate at 10 µg

Secondary
Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 110 kDa
Observed band size: 99 kDa why is the actual band size different from the predicted?
Additional bands at: 27 kDa, 56 kDa. We are unsure as to the identity of these extra bands.


Exposure time: 90 seconds


The 99 kDa band observed is comparable to the molecular weight seen with other commercially available antibodies to SERCA1 ATPase.

Anti-SERCA1 ATPase antibody [VE121G9] (ab2819) at 1/1000 dilution (in PBS tweeb 0.05% for 1 hour at 22°C) + Whole tissue lysate of human neck muscle. at 20 µg

Secondary
An HRP-conjugated sheep anti-mouse polyclonal at 1/4000 dilution

Developed using the ECL technique.

Predicted band size: 110 kDa
Observed band size: 110 kDa


Exposure time: 5 minutes


Blocking Step: 5% milk for 16 hours at 22°C

See Abreview

Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human heart 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 a dilution of 1:200 with a mouse monoclonal antibody recognizing SERCA1 ATPase ab2819 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 skeletal muscle 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 a dilution of 1:100 with a mouse monoclonal antibody recognizing SERCA1 ATPase ab2819 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.