Figure 1. Western blot analysis of Acid Phosphatase/ACP5 using anti-Acid Phosphatase/ACP5 antibody (MBS1755208).
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 HepG2 whole cell lysates,
Lane 2: human A431 whole cell lysates,
Lane 3: human Jurkat whole cell lysates,
Lane 4: human Hela whole cell lysates,
Lane 5: human K562 whole cell lysates,
Lane 6: human Raji whole cell lysates,
Lane 7: rat lung tissue lysates,
Lane 8: mouse lung 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-Acid Phosphatase/ACP5 antigen affinity purified polyclonal antibody (#MBS1755208) at 0.5 ug/mL overnight at 4 degree 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 with Tanon 5200 system. A specific band was detected for Acid Phosphatase/ACP5 at approximately 39 kDa. The expected band size for Acid Phosphatase/ACP5 is at 37 kDa.
Figure 2. IHC analysis of Acid Phosphatase/ACP5 using anti-Acid Phosphatase/ACP5 antibody (MBS1755208).
Acid Phosphatase/ACP5 was detected in a paraffin-embedded section of human spleen 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 2 ug/ml rabbit anti-Acid Phosphatase/ACP5 Antibody (MBS1755208) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
Figure 3. IHC analysis of Acid Phosphatase/ACP5 using anti-Acid Phosphatase/ACP5 antibody (MBS1755208).
Acid Phosphatase/ACP5 was detected in a paraffin-embedded section of mouse lung 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 2 ug/ml rabbit anti-Acid Phosphatase/ACP5 Antibody (MBS1755208) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
Figure 4. IHC analysis of Acid Phosphatase/ACP5 using anti-Acid Phosphatase/ACP5 antibody (MBS1755208).
Acid Phosphatase/ACP5 was detected in a paraffin-embedded section of human lung 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 2 ug/ml rabbit anti-Acid Phosphatase/ACP5 Antibody (MBS1755208) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
Figure 5. IHC analysis of Acid Phosphatase/ACP5 using anti-Acid Phosphatase/ACP5 antibody (MBS1755208).
Acid Phosphatase/ACP5 was detected in a paraffin-embedded section of human renal 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 2 ug/ml rabbit anti-Acid Phosphatase/ACP5 Antibody (MBS1755208) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.
Figure 6. IF analysis of Acid Phosphatase/ACP5 using anti-Acid Phosphatase/ACP5 antibody (MBS1755208).
Acid Phosphatase/ACP5 was detected in an immunocytochemical section of MCF-7 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 ug/mL rabbit anti-Acid Phosphatase/ACP5 Antibody (MBS1755208) overnight at 4 degree C. DyLight488 Conjugated Goat Anti-Rabbit IgG (MBS177405) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Figure 7. Flow Cytometry analysis of U251 cells using anti-Acid Phosphatase/ACP5 antibody (MBS1755208).
Overlay histogram showing U251 cells stained with MBS1755208 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Acid Phosphatase/ACP5 Antibody (MBS1755208, 1 ug/1x10^6 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG (MBS177405, 5-10 ug/1x10^6 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1 ug/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
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