MAG Antibody – Mouse Monoclonal

Figure 1. Western blot analysis of MAG using anti-MAG antibody (MBS1754017).
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: rat brain tissue lysates
Lane 2: mouse brain tissue lysates.
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- MAG antigen affinity purified monoclonal antibody (Catalog # MBS1754017) at 0. 25 μg/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 (Catalog # MBS176445) with Tanon 5200 system. A specific band was detected for MAG at approximately 100KD. The expected band size for MAG is at 100KD.

Figure 2. IHC analysis of MAG using anti-MAG antibody (MBS1754017).
MAG was detected in paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-MAG Antibody (MBS1754017) 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) (Catalog # MBS176438) with DAB as the chromogen.

Figure 3. IHC analysis of MAG using anti-MAG antibody (MBS1754017).
MAG was detected in paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-MAG Antibody (MBS1754017) 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) (Catalog # MBS176438) with DAB as the chromogen.

Figure 4. Flow Cytometry analysis of U87 cells using anti-MAG antibody (MBS1754017).
Overlay histogram showing U87 cells stained with MBS1754017 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti- MAG Antibody (MBS1754017, 1μg/1x10⁶ cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.