Human Osteopontin Recombinant Rabbit Monoclonal Antibody [PSH08-59]
cat.: HA723005
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: ELISA(Det)
Clonality: Monoclonal
Clone number: PSH08-59
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: PBS (pH7.4).
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Isotype: IgG
Immunogen: Recombinant protein within Human Osteopontin aa 17-314 (HA210993).
Positive control: Recombinant Human Osteopontin protein (HA210993).
Subcellular location: Secreted.
Recommended Dilutions:
  ELISA(Det)

Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Rabbit monoclonal [PSH08-58] to Human Osteopontin antibody (Capture) (HA723004) and recombinant Human Osteopontin protein (HA210993) as the standard. The reference range value is 61.7-5000 pg/ml.
Uniprot #: SwissProt: P10451 Human
Alternative names: BNSP Bone sialoprotein 1 Bone sialoprotein I BSP I BSPI Early T lymphocyte activation 1 ETA 1 ETA1 MGC110940 Nephropontin OPN Osteopontin osteopontin/immunoglobulin alpha 1 heavy chain constant region fusion protein OSTP_HUMAN PSEC0156 secreted phosphoprotein 1 (osteopontin bone sialoprotein I early T lymphocyte activation 1) secreted phosphoprotein 1 (osteopontin, bone sialoprotein I, early T-lymphocyte activation 1) Secreted phosphoprotein 1 SPP 1 SPP-1 SPP1 SPP1/CALPHA1 fusion Urinary stone protein Uropontin
Images
HA723005_1.png Fig1: Sandwich ELISA analysis of Human Osteopontin matched pair antibodies

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723004) diluted in carbonate/bicarbonate buffer, at a concentration of 5ug/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 Osteopontin protein (HA210993) starting from 5,000 pg/ml to 0 pg/ml and detect antibody (HA723005, 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.
HA723005_2.png Fig2: Interpolated concentrations of native Osteopontin in human samples.

Interpolated concentration of native Osteopontin was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean Osteopontin concentration was determined to be 609 ng/mL in human urine, 27 ng/ml in U-87 MG cell culture supernatant. There was no detectable signal in Daudi cell supernatant.
HA723005_3.png Fig3: Interpolated concentrations of spiked Osteopontin in cell culture media samples.

The concentrations of Osteopontin were measured in duplicates, interpolated from the Osteopontin 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”.