Human VEGFA Recombinant Rabbit Monoclonal Antibody [PSH12-91]
cat.: HA723493
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: ELISA(Cap)
Clonality: Monoclonal
Clone number: PSH12-91
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 VEGF aa 27-147 (HA210929).
Positive control: Recombinant Human VEGFA protein (HA210929).
Subcellular location: Secreted.
Recommended Dilutions:
  ELISA(Cap)
Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Rabbit monoclonal [PSH12-92] to Human VEGFA antibody (Detector) (HA723494) or Rabbit monoclonal [PSH12-93] to Human VEGFA antibody (Detector) (HA723496) and recombinant Human VEGFA protein (HA210929) as the standard. The reference range value is 31.3-4,000 pg/mL.
Uniprot #: SwissProt: P15692 Human
Alternative names: Folliculostellate cell-derived growth factor Glioma-derived endothelial cell mitogen MGC70609 MVCD1 Vascular endothelial growth factor A vascular endothelial growth factor A121 vascular endothelial growth factor A165 vascular endothelial growth factor Vascular permeability factor VEGF A Vegf VEGF-A VEGF120 Vegfa VEGFA_HUMAN VPF
Images
HA723493_1.png Fig1: Sandwich ELISA analysis of Human VEGFA matched pair antibodies

Capture: HA723493, Human VEGFA Rabbit mAb [PSH12-91]
Detector: HA723494, Human VEGFA Rabbit mAb [PSH12-92]

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA723493) diluted in carbonate/bicarbonate buffer, at a concentration of 5 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 VEGFA protein (HA210929) starting from 4,000 pg/ml to 0 pg/ml and detect antibody (HA723494, Biotin, 0.2 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.
HA723493_2.png Fig2: Sandwich ELISA analysis of Human VEGFA matched pair antibodies

Capture: HA723493, Human VEGFA Rabbit mAb [PSH12-91]
Detector: HA723496, Human VEGFA Rabbit mAb [PSH12-93]

Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody (HA723493) diluted in carbonate/bicarbonate buffer, at a concentration of 5 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 VEGFA protein (HA210929) starting from 4,000 pg/ml to 0 pg/ml and detect antibody (HA723496, Biotin, 0.2 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.
HA723493_3.png Fig3: Interpolated concentrations of native VEGFA in A431 and HepG2 cell culture supernatant.

Capture: HA723493, Human VEGFA Rabbit mAb [PSH12-91]
Detector: HA723494, Human VEGFA Rabbit mAb [PSH12-92]

The concentrations of VEGFA measured in duplicate and interpolated from the VEGFA standard curve and corrected for sample dilution. Undiluted samples are as follows: A431 cell culture supernatant 12.5% and HepG2 cell culture supernatant 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean VEGFA concentration was determined to be 6,033 pg/ml in A431 cell culture supernatant and 3,949 pg/ml in HepG2 cell culture supernatant.
HA723493_4.png Fig4: Interpolated concentrations of native VEGFA in A431 and HepG2 cell culture supernatant.

Capture: HA723493, Human VEGFA Rabbit mAb [PSH12-91]
Detector: HA723496, Human VEGFA Rabbit mAb [PSH12-93]

The concentrations of VEGFA measured in duplicate and interpolated from the VEGFA standard curve and corrected for sample dilution. Undiluted samples are as follows: A431 cell culture supernatant 12.5% and HepG2 cell culture supernatant 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean VEGFA concentration was determined to be 6,139 pg/ml in A431 cell culture supernatant and 3,773 pg/ml in HepG2 cell culture supernatant.
HA723493_5.png Fig5: Interpolated concentrations of spiked VEGFA in human cell culture media samples.

Capture: HA723493, Human VEGFA Rabbit mAb [PSH12-91]
Detector: HA723494, Human VEGFA Rabbit mAb [PSH12-92]

The concentrations of VEGFA were measured in duplicates, interpolated from the VEGFAstandard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).
HA723493_6.png Fig6: Interpolated concentrations of spiked VEGF in human cell culture media samples.

Capture: HA723493, Human VEGFA Rabbit mAb [PSH12-91]
Detector: HA723496, Human VEGFA Rabbit mAb [PSH12-93]

The concentrations of VEGFA were measured in duplicates, interpolated from the VEGFA standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 25%. 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”.