c-Kit Antibody

• Catalog Number: orb234304
• Category: Antibodies
• Description: c-Kit Antibody
• Clonality: Polyclonal
• Species/Host: Rabbit
• Isotype: Rabbit IgG
• Conjugation: Unconjugated
• Reactivity: Human
• Form/Appearance: Lyophilized
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Purification: Immunogen affinity purified.
• Immunogen: E.coli-derived human c-Kit recombinant protein (Position: Q26-S285). Human c-Kit shares 66% amino acid (aa) sequence identity with mouse c-Kit.
• UniProt ID: P10721
• MW: 109 kDa
• Tested applications: FC, ICC, IHC, WB
• Application notes: Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/mlImmunohistochemistry (Frozen Section), 0.5-1μg/ml Immunocytochemistry, 0.5-1μg/mlWestern blot, 0.1-0.5μg/mlFlow Cytometry (Fixed), 1-3μg/1x106cells. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
• Cross Reactivity: No cross-reactivity with other proteins
• Antibody Type: Primary Antibody
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Alternative names: Mast/stem cell growth factor receptor Kit; SCFR; 2
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Flow Cytometry analysis of K562 cells using anti-C-Kit antibody. Overlay histogram showing K562 cells (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 (1 µg/1x106 cells) for 30 min at 20°C. DyLight488 conjugated goat anti-rabbit IgG (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.

IHC analysis of C-Kit using anti-C-Kit antibody. 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 1 µg/ml rabbit anti-C-Kit Antibody 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) with DAB as the chromogen.

Western blot analysis of C-Kit using anti-C-Kit antibody. 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 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 with Tanon 5200 system. A specific band was detected for C-Kit at approximately 109 KD. The expected band size for C-Kit is at 109 KD.

Western blot analysis of C-Kit using anti-C-Kit antibody. 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 0.5 ng. 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 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 with Tanon 5200 system. A specific band was detected for C-Kit at approximately 49 KD. The expected band size for C-Kit is at 49 KD.