Anti-hnRNP K/HNRNPK Antibody Picoband®

Figure 1. Western blot analysis of hnRNP K/HNRNPK using anti-hnRNP K/HNRNPK antibody (A01793-2).
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 Hela whole cell lysates,
Lane 2: human MCF-7 whole cell lysates,
Lane 3: human MOLT-4 whole cell lysates,
Lane 4: human Daudi whole cell lysates,
Lane 5: human U251 whole cell lysates,
Lane 6: rat brain tissue lysates,
Lane 7: rat PC-12 whole cell lysates,
Lane 8: mouse brain tissue lysates,
Lane 9: 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-hnRNP K/HNRNPK antigen affinity purified polyclonal antibody (Catalog # A01793-2) at 0.5 μg/mL overnight at 4°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 # EK1002) with Tanon 5200 system. A specific band was detected for hnRNP K/HNRNPK at approximately 60 kDa. The expected band size for hnRNP K/HNRNPK is at 55 kDa.

Figure 2. IF analysis of hnRNP K/HNRNPK using anti-hnRNP K/HNRNPK antibody (A01793-2).
hnRNP K/HNRNPK was detected in an immunocytochemical section of U20S 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 μg/mL rabbit anti-hnRNP K/HNRNPK Antibody (A01793-2) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°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 HepG2 cells using anti-hnRNP K/HNRNPK antibody (A01793-2).
Overlay histogram showing HepG2 cells stained with A01793-2 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-hnRNP K/HNRNPK Antibody (A01793-2, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Figure 4. Flow Cytometry analysis of ANA-1 cells using anti-hnRNP K/HNRNPK antibody (A01793-2).
Overlay histogram showing ANA-1 cells stained with A01793-2 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-hnRNP K/HNRNPK Antibody (A01793-2, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Figure 5. Flow Cytometry analysis of C6 cells using anti-hnRNP K/HNRNPK antibody (A01793-2).
Overlay histogram showing C6 cells stained with A01793-2 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-hnRNP K/HNRNPK Antibody (A01793-2, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.