Figure 1. Western blot analysis of NFATC4 using anti-NFATC4 antibody (MBS1750778).
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 testis tissue lysates,
Lane 2: mouse testis tissue lysates,
Lane 3: human Hela whole cell lysates,
Lane 4: human placenta tissue lysates,
Lane 5: human A549 whole cell lysates,
Lane 6: human SK-OV-3 whole cell 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 rabbit anti-NFATC4 antigen affinity purified polyclonal antibody 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 1:10000 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 NFATC4 at approximately 95120KD. The expected band size for NFATC4 is at 95KD.
Figure 2. Flow Cytometry analysis of U20S cells using anti-NFATC4 antibody (MBS1750778).
Overlay histogram showing U20S cells stained with MBS1750778 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-NFATC4 Antibody (MBS1750778,1ug/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.
Figure 3. Flow Cytometry analysis of A431 cells using anti-NFATC4 antibody (MBS1750778).
Overlay histogram showing A431 cells stained with MBS1750778 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-NFATC4 Antibody (MBS1750778,1ug/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.
Figure 4. Flow Cytometry analysis of SiHa cells using anti-NFATC4 antibody (MBS1750778).
Overlay histogram showing SiHa cells stained with MBS1750778 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-NFATC4 Antibody (MBS1750778,1ug/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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