Anti-c-Kit Antibody Picoband®

Western blot analysis of C-Kit using anti-C-Kit antibody (PB9258).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours.
Lane 1: Recombinan Human C-Kit Protein 0.5ng
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-C-Kit antigen affinity purified polyclonal antibody (Catalog # PB9258) 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 1:10000 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 C-Kit at approximately 49KD. The expected band size for C-Kit is at 49KD.

Lumiflavin treatment reduces the development of DDP resistance of ovarian cancer OVCAR-3 cells line that is associated with cancer stem cells (CSCs). DDP resistance of ovarian cancer OVCAR-3 cell lines were induced by gradient concentration increment method ( ), and treated with 10, 20, 40, and 80 μm lumiflavin intervention simultaneously. The cell inhibition rate and resistance index of OVCAR-3cells to DDP were detected by CCK-8 assay kit. The proportion of CSCs (CD133+/CD177+ double positive cells) in OVCAR-3 cells was detected by flow cytometry. (A) Inhibitory curves of lumiflavin and DDP on OVCAR-3 and OVCAR-3/DDP cells. (B) Resistance index of OVCAR-3/DDP cells compared with OVCAR-3 cells. (C) Flow detection results of CD133+/CD117+ cells ratio of OVCAR-3 and OVCAR-3/DDP cells. (D) Statistical analysis graph of (C) . Mean ± SD ( n = 3). ** P < 0.01 difference between groups; * P < 0.05 difference between groups. DDP, cisplatin; OVCAR-3/DDP, DDP resistant OVCAR-3 cell lines; LUM, lumiflavin.
Index in PubMed under a CC BY license. PMID: 35669418

Effects of lumiflavin treatment on phenotypic differentiation of DDP-resistant cancer stem cells (CSCs/DDP). CSCs, CSCs/DDP were treated with 80 μM DDP, and CSCs/DDP were treated with 80 μM DDP and 20, 40, 80 μM lumiflavin for 72 h. Co-expression of CD133+/CD117+, CD44+/CD177, and CD44+/CD24– of CSCs/DDP were detected through flow cytometry. (A) Flow detection results of CD133+/CD117+, CD44+/CD177, and CD44+/CD24- cells of CSCs/DDP. (B) Statistical analysis graph of (A) . ** P < 0.01 between groups. Mean ± SD ( n = 3). CSCs/DDP, CSCs from DDP resistant OVCAR-3 cell lines; LUM, lumiflavin.
Index in PubMed under a CC BY license. PMID: 35669418

Western blot analysis of C-Kit using anti-C-Kit antibody (PB9258).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours.
Lane 1: HEPG2 Whole Cell Lysate
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-C-Kit antigen affinity purified polyclonal antibody (Catalog # PB9258) 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 1:10000 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 C-Kit at approximately 109KD. The expected band size for C-Kit is at 109KD.

IHC analysis of C-Kit using anti-C-Kit antibody (PB9258).
C-Kit was detected in paraffin-embedded section of human intestinal cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-C-Kit Antibody (PB9258) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

Flow Cytometry analysis of K562 cells using anti-C-Kit antibody (PB9258).
Overlay histogram showing K562 cells stained with PB9258 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-C-Kit Antibody (PB9258,1μg/1x106 cells) for 30 min at 20°C. DyLight488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.