SPAK/STK39 (STK39) Antibody – Rabbit Polyclonal

Figure 1. IF analysis of SPAK/STK39 using anti- SPAK/STK39 antibody (MBS1753532).
SPAK/STK39 was detected in immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti- SPAK/STK39 Antibody (MBS1753532) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Rabbit IgG 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 2. Western blot analysis of SPAK/STK39 using anti-SPAK/STK39 antibody (MBS1753532).
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: human DAUDI whole cell lysates
Lane 2: human HL-60 whole cell lysates
Lane 3: rat brain tissue lysates
Lane 4: rat testis tissue lysates
Lane 5: mouse brain tissue lysates
Lane 6: mouse testis tissue 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-SPAK/STK39 antigen affinity purified polyclonal antibody (Catalog # MBS1753532) at 0. 5 μg/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 (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for SPAK/STK39 at approximately 65-70KD. The expected band size for SPAK/STK39 is at 65-70KD.

Figure 3. Flow Cytometry analysis of HL-60 cells using anti-SPAK/STK39 antibody (MBS1753532).
Overlay histogram showing HL-60 cells stained with MBS1753532 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-SPAK/STK39 Antibody (MBS1753532, 1μg/1x10⁶ cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.