HA723872

P-Selectin / CD62P Recombinant Rabbit Monoclonal Antibody [PSH16-70]

cat.: HA723872

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	ELISA(Cap)
Clonality:	Monoclonal
Clone number:	PSH16-70

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 Mouse P-Selectin / CD62P aa 42-709 (HA211140).
Positive control:	Recombinant Human P-Selectin / CD62P protein (HA210931) or Recombinant Mouse P-Selectin / CD62P protein (HA211140).
Subcellular location:	Cell membrane.
Recommended Dilutions: ELISA(Cap)	Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Rabbit monoclonal [PSH16-71] to P-Selectin / CD62P antibody (Detector) (HA723873) and Recombinant Human P-Selectin / CD62P protein (HA210931) or Recombinant Mouse P-Selectin / CD62P protein (HA211140) as the standard. The reference range value is 78.1-20,000 pg/mL.
Uniprot #:	SwissProt: P16109 Human \| Q01102 Mouse \| P98106 Rat
Alternative names:	Antigen CD62 CD62 CD62 antigen-like family member P CD62P GMP 140 GMP-140 GMP140 GMRP Granule membrane protein 140 Granulocyte membrane protein GRMP LECAM 3 LECAM3 Leukocyte endothelial cell adhesion molecule 3 Leukocyte-endothelial cell adhesion molecule 3 LYAM3_HUMAN P Selectin P-selectin PADGEM Platelet activation dependent granule-external membrane protein Platelet alpha granule membrane protein PSEL Selectin P (granule membrane protein 140kDa, antigen CD62) SELP

Images

Fig1: Sandwich ELISA analysis of Human P-Selectin matched pair antibodies

Capture: HA723872, P-Selectin / CD62P Rabbit mAb [PSH16-70]
Detector: HA723873, P-Selectin / CD62P Rabbit mAb [PSH16-71]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723872) 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 P-Selectin / CD62P protein (HA210931) starting from 20,000 pg/ml to 0 pg/ml and detect antibody (HA723873, 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 mouse P-Selectin matched pair antibodies

Capture: HA723872, P-Selectin / CD62P Rabbit mAb [PSH16-70]
Detector: HA723873, P-Selectin / CD62P Rabbit mAb [PSH16-71]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723872) 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 Mouse P-Selectin / CD62P protein (HA211140) starting from 20,000 pg/ml to 0 pg/ml and detect antibody (HA723873, 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 P-Selectin in human serum samples and HeLa cell culture supernatant. Capture: HA723872, P-Selectin / CD62P Rabbit mAb [PSH16-70] Detector: HA723873, P-Selectin / CD62P Rabbit mAb [PSH16-71] The concentrations of P-Selectin were measured in triplicates, interpolated from the P-Selectin standard curve and corrected for sample dilution. Undiluted samples are human serum 5% and HeLa cell culture supernatant 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=3). The mean P-Selectin concentration was determined to be 95,663 pg/ml in human serum and undetectable in HeLa cell culture supernatant.
	Fig4: Interpolated concentrations of native P-Selectin in mouse serum samples. Capture: HA723872, P-Selectin / CD62P Rabbit mAb [PSH16-70] Detector: HA723873, P-Selectin / CD62P Rabbit mAb [PSH16-71] The concentrations of P-Selectin were measured in duplicates, interpolated from the P-Selectin standard curve and corrected for sample dilution. Undiluted samples are mouse serum 1.25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean P-Selectin concentration was determined to be 849,328 pg/ml in mouse serum.
	Fig5: Interpolated concentrations of native P-Selectin in rat serum samples. Capture: HA723872, P-Selectin / CD62P Rabbit mAb [PSH16-70] Detector: HA723873, P-Selectin / CD62P Rabbit mAb [PSH16-71] The concentrations of P-Selectin were measured in duplicates, interpolated from the P-Selectin standard curve and corrected for sample dilution. Undiluted samples are rat serum 1.25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean P-Selectin concentration was determined to be 118,074 pg/ml in rat serum.

Fig6: Interpolated concentrations of spiked P-Selectin in human cell culture media samples.

Capture: HA723872, P-Selectin / CD62P Rabbit mAb [PSH16-70]
Detector: HA723873, P-Selectin / CD62P Rabbit mAb [PSH16-71]

The concentrations of P-Selectin were measured in duplicates, interpolated from the P-Selectin 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).

Fig7: Interpolated concentrations of spiked P-Selectin in mouse cell culture media samples.

Capture: HA723872, P-Selectin / CD62P Rabbit mAb [PSH16-70]
Detector: HA723873, P-Selectin / CD62P Rabbit mAb [PSH16-71]

The concentrations of P-Selectin were measured in duplicates, interpolated from the P-Selectin 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”.