Biotin Conjugated Human Nectin-2 Recombinant Rabbit Monoclonal Antibody [PSH12-52]
cat.: HA723462B
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: ELISA(Det), ELISA
Clonality: Monoclonal
Clone number: PSH12-52
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% ProClin300.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Isotype: IgG
Immunogen: Recombinant protein within Human Nectin-2 aa 32-360 (HA210905).
Positive control: Recombinant Human Nectin-2 protein (HA210905).
Subcellular location: Cell membrane.
Recommended Dilutions:
  ELISA(Det)
  ELISA
Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Rabbit monoclonal [PSH11-54] to Human Nectin-2 antibody (Capture) (HA723460) or Rabbit monoclonal [PSH12-53] to Human Nectin-2 antibody (Capture) (HA723463) and Recombinant Human Nectin-2 protein (HA210905) as the standard. The reference range value is 39.1-5,000 pg/mL.
Use at an assay dependent concentration.
Uniprot #: SwissProt: Q92692 Human
Alternative names: CD 112 CD112 CD112 antigen Herpes virus entry mediator B Herpes virus entry protein B Herpesvirus entry mediator B Herpesvirus entry protein B Hve B HveB Nectin-2 Nectin2 Poliovirus receptor like 2 Poliovirus receptor related 2 (herpesvirus entry mediator B) Poliovirus receptor related 2 Poliovirus receptor related protein 2 Poliovirus receptor-related protein 2 PRR 2 PRR2 PVRL 2 PVRL2 PVRL2_HUMAN PVRR 2 PVRR2
Images
HA723462B_1.png Fig1: Sandwich ELISA analysis of human Nectin-2 matched pair antibodies

Capture: HA723460, Human Nectin-2 Rabbit mAb [PSH11-54]
Detector: HA723461, Human Nectin-2 Rabbit mAb [PSH12-52]

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA723460) diluted in carbonate/bicarbonate buffer, at a concentration of 1 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 Nectin-2 protein (HA210905) starting from 5,000 pg/ml to 0 pg/ml and detect antibody (HA723461, Biotin, 0.05 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 50 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.
HA723462B_2.png Fig2: Sandwich ELISA analysis of human Nectin-2 matched pair antibodies

Capture: HA723463, Human Nectin-2 Rabbit mAb [PSH12-53]
Detector: HA723461, Human Nectin-2 Rabbit mAb [PSH12-52]

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA723463) diluted in carbonate/bicarbonate buffer, at a concentration of 1 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 Nectin-2 protein (HA210905) starting from 5,000 pg/ml to 0 pg/ml and detect antibody (HA723461, Biotin, 0.05 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 50 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.
HA723462B_3.png Fig3: Interpolated concentrations of native Nectin-2 in MCF7 and MOLT-4 extract samples based on a 1,000 µg/ml extract load.

Capture: HA723460, Human Nectin-2 Rabbit mAb [PSH11-54]
Detector: HA723461, Human Nectin-2 Rabbit mAb [PSH12-52]

The concentrations of Nectin-2 were measured in duplicates, interpolated from the Nectin-2 standard curve and corrected for sample dilution. Undiluted samples are MOLT-4 extract 50% and MCF7 extract 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Nectin-2 concentration was determined to be 3,548 pg/ml in MCF7 extract and undetectable in MOLT-4 extract.
HA723462B_4.png Fig4: Interpolated concentrations of native Nectin-2 in MCF7 and MOLT-4 extract samples based on a 1,000 µg/ml extract load.

Capture: HA723463, Human Nectin-2 Rabbit mAb [PSH12-53]
Detector: HA723461, Human Nectin-2 Rabbit mAb [PSH12-52]

The concentrations of Nectin-2 were measured in duplicates, interpolated from the Nectin-2 standard curve and corrected for sample dilution. Undiluted samples are MOLT-4 extract 50% and MCF7 extract 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Nectin-2 concentration was determined to be 4,062 pg/ml in MCF7 extract and undetectable in MOLT-4 extract.
HA723462B_5.png Fig5: Interpolated concentrations of spiked Nectin-2 in human cell culture media samples.

Capture: HA723460, Human Nectin-2 Rabbit mAb [PSH11-54]
Detector: HA723461, Human Nectin-2 Rabbit mAb [PSH12-52]

The concentrations of Nectin-2 were measured in duplicates, interpolated from the Nectin-2 standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).
HA723462B_6.png Fig6: Interpolated concentrations of spiked Nectin-2 in human cell culture media samples.

Capture: HA723463, Human Nectin-2 Rabbit mAb [PSH12-53]
Detector: HA723461, Human Nectin-2 Rabbit mAb [PSH12-52]

The concentrations of Nectin-2 were measured in duplicates, interpolated from the Nectin-2 standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 50%. 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”.