Figure 1. IHC analysis of CD20 using anti-CD20 antibody (MBS176945).
CD20 was detected in paraffin-embedded section of Human Tonsil 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 1ug/ml rabbit anti-CD20 Antibody (MBS176945) 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 2. IHC analysis of CD20 using anti-CD20 antibody (MBS176945).
CD20 was detected in paraffin-embedded section of Human Tonsil 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 1ug/ml rabbit anti-CD20 Antibody (MBS176945) 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. Western blot analysis of CD20 using anti-CD20 antibody (MBS176945).
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: RAJI Whole Cell Lysate
Lane 2: CEM Whole Cell 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 rabbit anti-CD20 antigen affinity purified polyclonal antibody at 0.5ug/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 1:10000 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 CD20 at approximately 33KD. The expected band size for CD20 is at 33KD.
Figure 4. Flow Cytometry analysis of U937 cells using anti-CD20 antibody (MBS176945).
Overlay histogram showing U937 cells stained with MBS176945 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CD20 Antibody (MBS176945,1ug/1x10^6 cells) for 30 min at 20 degree C. DyLight 488 conjugated goat anti-rabbit IgG (5-10ug/1x10^6 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
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