Figure 1. Western blot analysis of RAD21 using anti-RAD21 antibody (MBS1754999).
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: monkey COS-7 whole cell lysates,
Lane 2: human Hela whole cell lysates,
Lane 3: human MOLT-4 whole cell lysates,
Lane 4: human Jurkat whole cell lysates,
Lane 5: human HEL whole cell lysates,
Lane 6: human Daudi whole cell lysates,
Lane 7: human A431 whole cell lysates,
Lane 8: human T-47D whole cell lysates,
Lane 9: rat brain tissue lysates,
Lane 10: rat pancreas tissue lysates,
Lane 11: mouse brain tissue lysates,
Lane 12: mouse pancreas 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-RAD21 antigen affinity purified polyclonal antibody (#MBS1754999) 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 RAD21 at approximately 72 kDa. The expected band size for RAD21 is at 72 kDa.
Figure 2. IF analysis of RAD21 using anti-RAD21 antibody (MBS1754999).
RAD21 was detected in an immunocytochemical section of SiHa 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-RAD21 Antibody (MBS1754999) 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 HL-60 cells using anti-RAD21 antibody (MBS1754999).
Overlay histogram showing HL-60 cells stained with MBS1754999 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-RAD21 Antibody (MBS1754999, 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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