Figure 1. Western blot analysis of TRIP/TRAIP using anti-TRIP/TRAIP antibody (MBS1755626).
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 U251 whole cell lysates,
Lane 3: rat brain tissue lysates,
Lane 4: mouse brain tissue 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-TRIP/TRAIP antigen affinity purified polyclonal antibody (#MBS1755626) 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 TRIP/TRAIP at approximately 53 kDa. The expected band size for TRIP/TRAIP is at 53 kDa.
Figure 2. IF analysis of TRIP/TRAIP using anti-TRIP/TRAIP antibody (MBS1755626).
TRIP/TRAIP was detected in an immunocytochemical section of HeLa 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-TRIP/TRAIP Antibody (MBS1755626) 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 3. Flow Cytometry analysis of SiHa cells using anti-TRIP/TRAIP antibody (MBS1755626).
Overlay histogram showing SiHa cells stained with MBS1755626 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-TRIP/TRAIP Antibody (MBS1755626, 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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