Anti-GRP94/HSP90B1 Antibody Picoband®

Figure 1. Western blot analysis of GRP94/HSP90B1 using anti-GRP94/HSP90B1 antibody (PA1340).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human A549 whole cell lysates,
Lane 2: human Hela whole cell lysates,
Lane 3: human Jurkat whole cell lysates,
Lane 4: human K562 whole cell 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-GRP94/HSP90B1 antigen affinity purified polyclonal antibody (Catalog # PA1340) 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 Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for GRP94/HSP90B1 at approximately 100 kDa. The expected band size for GRP94/HSP90B1 is at 92 kDa.

Figure 2. ICC analysis of GRP94/HSP90B1 using anti-GRP94/HSP90B1 antibody (PA1340).
GRP94/HSP90B1 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 1 μg/ml rabbit anti-GRP94/HSP90B1 Antibody (PA1340) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

Figure 3. IHC analysis of GRP94/HSP90B1 using anti-GRP94/HSP90B1 antibody (PA1340).
GRP94/HSP90B1 was detected in a paraffin-embedded section of Human Lung Cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-GRP94/HSP90B1 Antibody (PA1340) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

Figure 4. IHC analysis of GRP94/HSP90B1 using anti-GRP94/HSP90B1 antibody (PA1340).
GRP94/HSP90B1 was detected in a paraffin-embedded section of Rat Brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-GRP94/HSP90B1 Antibody (PA1340) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

Figure 4. Western blot analysis of GRP94/HSP90B1 using anti-GRP94/HSP90B1 antibody (PA1340).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% BSA for 1.5 hour at RT. The membrane was incubated with rabbit anti-GRP94/HSP90B1 antibody (PA1340) at 1:2000 in TBST/5% BSA 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 Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for GRP94/HSP90B1 at approximately 92 kDa. The expected band size for GRP94/HSP90B1 is at 92 kDa.