Figure 1. Western blot analysis of WIPF1 using anti-WIPF1 antibody (MBS1755368).
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 Daudi whole cell lysates,
Lane 2: human MOLT-4 whole cell lysates,
Lane 3: rat spleen tissue lysates,
Lane 4: mouse spleen tissue lysates,
Lane 5: 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-WIPF1 antigen affinity purified polyclonal antibody (#MBS1755368) 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 WIPF1 at approximately 56 kDa. The expected band size for WIPF1 is at 51 kDa.
Figure 2. Flow Cytometry analysis of HEL cells using anti-WIPF1 antibody (MBS1755368).
Overlay histogram showing HEL cells stained with MBS1755368 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-WIPF1 Antibody (MBS1755368, 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.
Figure 3. Flow Cytometry analysis of U87 cells using anti-WIPF1 antibody (MBS1755368).
Overlay histogram showing U87 cells stained with MBS1755368 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-WIPF1 Antibody (MBS1755368, 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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