c-Kit Recombinant Rabbit Monoclonal Antibody [PD00-24]
cat.: HA721154
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: IHC-P, WB, mIHC
Clonality: Monoclonal
Clone number: PD00-24
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 110 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human c-Kit aa 950-976.
Positive control: Human seminoma tissue, human gastrointestinal stromal tumor tissue, human breast tissue, Saos-2 cell lysate, human lung tissue lysate, human cervical cancer.
Subcellular location: Cell membrane; Cytoplasm.
Recommended Dilutions:
  IHC-P
  WB
  mIHC
1:1,000
1:1,000
1:1,000
Uniprot #: SwissProt: P10721 Human
Alternative names: C Kit c-Kit c-Kit Ligand CD117 Kit Kit Ligand KIT oncogene KIT proto oncogene receptor tyrosine kinase KIT_HUMAN Mast cell growth factor receptor Mast/stem cell growth factor receptor Kit MGF p145 c-kit PBT Piebald trait protein Proto oncogene c Kit Proto oncogene tyrosine protein kinase Kit Proto-oncogene c-Kit SCF Receptor SCFR soluble KIT variant 1 Steel Factor Receptor Stem cell factor receptor tyrosine protein kinase Kit Tyrosine-protein kinase Kit v kit Hardy Zuckerman 4 feline sarcoma viral oncogene homolog v kit Hardy Zuckerman 4 feline sarcoma viral oncogene like protein v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog
Images
HA721154_1.jpg Fig1: Fluorescence multiplex immunohistochemical analysis of the human cervical cancer (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-CD57 (HA601114, red), anti-CD11c (ET1606-19, green), anti-CD117 (HA21154, magenta) and anti-CD66b (HA500100, yellow) on human cervical cancer. Panel B: anti- CD57 stained on NKT cells. Panel C: anti-CD11c stained on dendritic cells. Panel D: anti-CD117 stained on mast cells. Panel E: anti-CD66b stained on neutrophils. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKit™MH010101, www.luminiris.cn). The section was incubated in four rounds of staining: in the order of HA601114 (1/500 dilution), ET1606-19 (1/1,000 dilution), HA721154 (1/1,000 dilution), and HA500100 (1/1,000 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.
HA721154_2.jpg Fig2: Fluorescence multiplex immunohistochemical analysis of human cervical carcinoma (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-S100A9 (ET1702-73, White), anti-CD117 (HA721154, Red) and anti-CD163(ET1704-43, Yellow) on human cervical carcinoma. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKit™MH010101, www.luminiris.cn). The section was incubated in three rounds of staining: in the order of ET1702-73 (1/1,000 dilution), HA721154 (1/1,000 dilution) and ET1704-43 (1/2,000 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Zeiss Observer 7 Inverted Fluorescence Microscope.
HA721154_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human seminoma tissue with Rabbit anti-c-Kit antibody (HA721154) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721154) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA721154_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human gastrointestinal stromal tumor tissue with Rabbit anti-c-Kit antibody (HA721154) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721154) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA721154_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human breast tissue with Rabbit anti-c-Kit antibody (HA721154) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721154) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA721154_6.jpg Fig6: Western blot analysis of c-Kit on different lysates with Rabbit anti-c-Kit antibody (HA721154) at 1/1,000 dilution.

Lane 1: Saos-2 cell lysate
Lane 2: HL-60 cell lysate (low expression)
Lane 3: Jurkat cell lysate (low expression)
Lane 4: Human lung tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 110 kDa
Observed band size: 150 kDa

Exposure time: 2 minutes;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721154) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.
HA721154_7.jpg Fig7: Western blot analysis of c-Kit on different lysates with Rabbit anti-c-Kit antibody (HA721154) at 1/1,000 dilution.

Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-c-Kit KD cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 110 kDa
Observed band size: 150 kDa

Exposure time: 2 minutes 40 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721154) at 1/1,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.