Figure 1. Flow Cytometry analysis of U20S cells using anti-GSTM1 antibody (MBS1751984).
Overlay histogram showing U20S cells stained with MBS1751984 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-GSTM1 Antibody (MBS1751984, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-mouse IgG (BA1126, 5-10ug/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Figure 3. IHC analysis of GSTM1 using anti-GSTM1 antibody (MBS1751984).
GSTM1 was detected in paraffin-embedded section of rat liver tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml mouse anti-GSTM1 Antibody (MBS1751984) overnight at 4 degree C. Biotinylated goat anti-mouse 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 GSTM1 using anti-GSTM1 antibody (MBS1751984).
GSTM1 was detected in paraffin-embedded section of rat liver tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml mouse anti-GSTM1 Antibody (MBS1751984) overnight at 4 degree C. Biotinylated goat anti-mouse 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 GSTM1 using anti-GSTM1 antibody (MBS1751984).
GSTM1 was detected in paraffin-embedded section of human colon cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml mouse anti-GSTM1 Antibody (MBS1751984) overnight at 4 degree C. Biotinylated goat anti-mouse 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. Western blot analysis of GSTM1 using anti-GSTM1 antibody (MBS1751984).
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 50ug of sample under reducing conditions.
Lane 1: human Hela, whole cell lysate,
Lane 2: human T-47D whole cell lysate,
Lane 3: rat brain tissue lysate,
Lane 4: rat lung tissue lysate,
Lane 5: rat stomach tissue lysate,
Lane 6: mouse lung tissue lysate,
Lane 7: mouse stomach tissue lysate,
Lane 8: mouse kidney tissue lysate.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-GSTM1 antigen affinity purified monoclonal antibody 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- mouse IgG-HRP secondary antibody at a dilution of 1:10000 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 GSTM1 at approximately 26KD. The expected band size for GSTM1 is at 26KD.
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