HA724619

Human c-Kit Recombinant Rabbit Monoclonal Antibody [PSH26-40]

cat.: HA724619

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human
Applications:	ELISA(Cap)
Clonality:	Monoclonal
Clone number:	PSH26-40

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*PBS (pH7.4).
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Isotype:	IgG
Immunogen:	Recombinant protein within Human c-Kit aa 26-516 (HA211233).
Positive control:	Recombinant Human c-Kit protein (HA211233).
Subcellular location:	Cell membrane; Cytoplasm.
Recommended Dilutions: ELISA(Cap)	Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Rabbit monoclonal [PSH26-41] to Human c-Kit (Detecor) (HA724620) or Rabbit monoclonal [PSH26-42] to Human c-Kit (Detecor) (HA724621) and Recombinant Human c-Kit protein (HA211233) as the standard. The reference range value is 11.7-3,000 pg/mL.
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

Fig1: Sandwich ELISA analysis of Human c-Kit matched pair antibodies

Capture: HA724619, Human c-Kit Rabbit mAb [PSH26-40]
Detector: HA724620, Human c-Kit Rabbit mAb [PSH26-41]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724619) diluted in carbonate/bicarbonate buffer, at a concentration of 2 μg/mL overnight at 4℃. Wells of the plate were washed, blocked with 150 µl 0.05% tween-20 1% BSA blocking buffer, and incubated with serial diluted Recombinant Human c-Kit protein (HA211233) starting from 3,000 pg/mL to 0 pg/mL and detect antibody (HA724620, Biotin, 0.2 μg/mL) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 100 µl per well of SA-HRP for 0.5 hour at 30℃ with shaking. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.

Fig2: Sandwich ELISA analysis of Human c-Kit matched pair antibodies

Capture: HA724619, Human c-Kit Rabbit mAb [PSH26-40]
Detector: HA724621, Human c-Kit Rabbit mAb [PSH26-42]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724619) diluted in carbonate/bicarbonate buffer, at a concentration of 2 μg/mL overnight at 4℃. Wells of the plate were washed, blocked with 150 µl 0.05% tween-20 1% BSA blocking buffer, and incubated with serial diluted Recombinant Human c-Kit protein (HA211233) starting from 3,000 pg/mL to 0 pg/mL and detect antibody (HA724621, Biotin, 0.2 μg/mL) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 100 µl per well of SA-HRP for 0.5 hour at 30℃ with shaking. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.

	Fig3: Interpolated concentrations of native c-Kit in IMR-32 cell extract samples based on a 1,000 µg/mL extract load. Capture: HA724619, Human c-Kit Rabbit mAb [PSH26-40] Detector: HA724620, Human c-Kit Rabbit mAb [PSH26-41] Interpolated concentration of native c-Kit was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean c-Kit concentration was determined to be 4,666 pg/mL in IMR-32 cell extract.
	Fig4: Interpolated concentrations of native c-Kit in IMR-32 cell extract samples based on a 1,000 µg/mL extract load. Capture: HA724619, Human c-Kit Rabbit mAb [PSH26-40] Detector: HA724621, Human c-Kit Rabbit mAb [PSH26-42] Interpolated concentration of native c-Kit was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean c-Kit concentration was determined to be 7,935 pg/mL in IMR-32 cell extract.
	Fig5: Interpolated concentrations of native c-Kit in human serum. Capture: HA724619, Human c-Kit Rabbit mAb [PSH26-40] Detector: HA724620, Human c-Kit Rabbit mAb [PSH26-41] Interpolated concentration of native c-Kit was measured in duplicate at different sample concentrations and interpolated from the c-Kit standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean c-Kit concentration was determined to be 116,136 pg/mL in human serum.
	Fig6: Interpolated concentrations of native c-Kit in human serum. Capture: HA724619, Human c-Kit Rabbit mAb [PSH26-40] Detector: HA724621, Human c-Kit Rabbit mAb [PSH26-42] Interpolated concentration of native c-Kit was measured in duplicate at different sample concentrations and interpolated from the c-Kit standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean c-Kit concentration was determined to be 163,987 pg/mL in human serum.

Fig7: Interpolated concentrations of spiked c-Kit in cell culture media samples.

Capture: HA724619, Human c-Kit Rabbit mAb [PSH26-40]
Detector: HA724620, Human c-Kit Rabbit mAb [PSH26-41]

The concentrations of c-Kit were measured in duplicates, interpolated from the c-Kit standard curves and corrected for sample dilution. diluted samples are as follows: 50% cell culture media with FBS. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

Fig8: Interpolated concentrations of spiked c-Kit in cell culture media samples.

Capture: HA724619, Human c-Kit Rabbit mAb [PSH26-40]
Detector: HA724621, Human c-Kit Rabbit mAb [PSH26-42]

The concentrations of c-Kit were measured in duplicates, interpolated from the c-Kit standard curves and corrected for sample dilution. diluted samples are as follows: 50% cell culture media with FBS. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.