Anti-GPSM1 Antibody Picoband®

Western blot analysis of GPSM1 using anti-GPSM1 antibody (A04971-1).
Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human SH-SY5Y whole cell lysates,
Lane 2: human 293T whole cell lysates,
Lane 3: human Hela whole cell lysates,
Lane 4: human U2OS whole cell lysates,
Lane 5: rat heart tissue lysates,
Lane 6: rat brain tissue lysates,
Lane 7: mouse heart tissue lysates,
Lane 8: mouse brain tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-GPSM1 antigen affinity purified polyclonal antibody (A04971-1) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for GPSM1 at approximately 80 kDa. The expected band size for GPSM1 is at 75 kDa.

IF analysis of GPSM1 using anti-GPSM1 antibody (A04971-1).
GPSM1 was detected in an immunocytochemical section of HELA cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-GPSM1 Antibody (A04971-1) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Immunoprecipitating GPSM1 in SH-SY5Y whole cell lysate.
Western blot analysis of GPSM1 using anti-GPSM1 antibody (A04971-1).
Lane 1: SH-SY5Y whole cell lysates (30ug),
Lane 2: Rabbit control IgG instead of anti-GPSM1 antibody in SH-SY5Y whole cell lysate,
Lane 3: anti-GPSM1 antibody (2μg) + SH-SY5Y whole cell lysate (500μg).
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-GPSM1 antigen affinity purified polyclonal antibody (A04971-1) at a dilution of 0.5 μg/mL and probed with a goat anti-rabbit IgG-HRP secondary antibody (Catalog # BA1054). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1197). A specific band was detected for GPSM1 at approximately 80 kDa. The expected band size for GPSM1 is at 75 kDa.

Flow Cytometry analysis of 293T cells using anti-GPSM1 antibody (A04971-1).
Overlay histogram showing 293T cells stained with A04971-1 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-GPSM1 Antibody (A04971-1, 1 μg/1x10⁶ cells) for 30 min at 20°C. Fluoro488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.