Figure 1. Western blot analysis of AHA1/AHSA1 using anti-AHA1/AHSA1 antibody (MBS1755486).
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 Daudi whole cell lysates,
Lane 3: human HepG2 whole cell lysates,
Lane 4: human Caco-2 whole cell lysates,
Lane 5: human MOLT4 whole cell lysates,
Lane 6: human Jurkat whole cell lysates,
Lane 7: rat testis tissue lysates,
Lane 8: rat brain tissue lysates,
Lane 9: rat kidney tissue lysates,
Lane 10: rat C6 whole cell lysates,
Lane 11: mouse brain tissue lysates,
Lane 12: mouse kidney tissue lysates,
Lane 13: mouse NIH/3T3 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-AHA1/AHSA1 antigen affinity purified polyclonal antibody (#MBS1755486) at 0.25 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 AHA1/AHSA1 at approximately 40 kDa. The expected band size for AHA1/AHSA1 is at 40 kDa.
Figure 2. IHC analysis of AHA1/AHSA1 using anti-AHA1/AHSA1 antibody (MBS1755486).
AHA1/AHSA1 was detected in a paraffin-embedded section of human breast infiltrating ductal carcinoma 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-AHA1/AHSA1 Antibody (MBS1755486) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.
Figure 3. IHC analysis of AHA1/AHSA1 using anti-AHA1/AHSA1 antibody (MBS1755486).
AHA1/AHSA1 was detected in a paraffin-embedded section of human colorectal adenocarcinoma 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-AHA1/AHSA1 Antibody (MBS1755486) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.
Figure 4. IHC analysis of AHA1/AHSA1 using anti-AHA1/AHSA1 antibody (MBS1755486).
AHA1/AHSA1 was detected in a paraffin-embedded section of human endometrial 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-AHA1/AHSA1 Antibody (MBS1755486) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.
Figure 5. IHC analysis of AHA1/AHSA1 using anti-AHA1/AHSA1 antibody (MBS1755486).
AHA1/AHSA1 was detected in a paraffin-embedded section of human laryngeal squamous cell carcinoma 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-AHA1/AHSA1 Antibody (MBS1755486) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.
Figure 6. IHC analysis of AHA1/AHSA1 using anti-AHA1/AHSA1 antibody (MBS1755486).
AHA1/AHSA1 was detected in a paraffin-embedded section of human liver 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-AHA1/AHSA1 Antibody (MBS1755486) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.
Figure 7. IHC analysis of AHA1/AHSA1 using anti-AHA1/AHSA1 antibody (MBS1755486).
AHA1/AHSA1 was detected in a paraffin-embedded section of mouse testis 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-AHA1/AHSA1 Antibody (MBS1755486) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.
Figure 8. IHC analysis of AHA1/AHSA1 using anti-AHA1/AHSA1 antibody (MBS1755486).
AHA1/AHSA1 was detected in a paraffin-embedded section of rat testis 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-AHA1/AHSA1 Antibody (MBS1755486) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.
Figure 9. IF analysis of AHA1/AHSA1 using anti-AHA1/AHSA1 antibody (MBS1755486).
AHA1/AHSA1 was detected in an immunocytochemical section of A431 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-AHA1/AHSA1 Antibody (MBS1755486) 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 10. Flow Cytometry analysis of HepG2 cells using anti-AHA1/AHSA1 antibody (MBS1755486).
Overlay histogram showing HepG2 cells stained with MBS1755486 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-AHA1/AHSA1 Antibody (MBS1755486, 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.