Human VEGF ELISA Kit, Lot 21AO-939 [Cancer Immune Checkpoint Assay Kit] (CAT#: IOK-05-P850)

Microstrips Coated w / Capture Antibody: 12 x 8-Well Microstrips.
Protein Standard: Lyophilized (100 ng), Red container.
Biotinylated Detection Antibody: Lyophilized, Yellow container.
400x Streptavidin-HRP: 30 μL, Blue container.
Wash Buffer (10x): 50 mL, Clear containter.
Assay Diluent: 50 mL, Clear container.
Ready-to-Use Substrate: 12 mL, Brown container.
Stop Solution: 12 mL, Clear container.
Adhesive Plate Sealers: 4 Sheets.
Technical Manual 1 Manual.

Microplate reader capable of measuring absorbance at 450nm, with the correction wavelength set at 540nm or 570nm.
An incubator which can provide stable incubation conditions up to 37°C ± 0.5°C.
Squirt bottle, manifold dispenser or automated microplate washer.
Absorbent paper for blotting the microtiter plate.
100mL and 500mL graduated cylinders.
Deionized or distilled water.
Pipettes and pipette tips.
Test tubes for dilution.

Pre-coated

1.Reconstitute the Biotin-Conjugated Detection Antibody in 67 µL of ddH₂O for a concentration of 180 µg/ml.
2.Reconstitute the Protein Standard in 100 µL of ddH₂O for a concentration of 340 ng/ml.
3.Dilute the 50 mL of 10x Wash Buffer in 450 mL of ddH2O for 500 mL of 1x Wash Buffer.

1.Average the duplicate or triplicate readings for each standard, control and sample and subtract the average zero standard optical density.
2.Generate a standard curve by using Microsoft Excel or other computer software capable of establishing a 4- Parameter Logistic (4-PL) curve fit. If using Excel or an alternative graphing tool, plot the average optical density values in absorbance units (y-axis) against the known standard concentrations in pg/ml (x-axis).

Reagents provided in this kit may be harmful if ingested, inhaled or absorbed through the skin. Please carefully review the MSDS for each reagent before conducting the experiment.
Stop Solution contains 2 N Sulfuric Acid (H2SO4) and is an extremely corrosive agent. Please wear proper eye, hand and face protection when handling this material. When the experiment is finished, be sure to rinse the plate with copious amounts of running water to dilute the Stop Solution prior to disposing the plate.

The kit should not be used beyond the expiration date on the kit label.
Do not mix or substitute reagents with those from other lots or sources.
If samples generate values higher than the highest standard, dilute the samples with Sample Diluent and repeat the assay.
Any variation in Sample Diluent, operator, pipetting technique, washing technique, incubation time/temperature and kit age can cause variation in binding.
This assay is designed to eliminate interference by soluble receptors, binding proteins and other factors present in biological samples. Until all factors have been tested in the Immunoassay, the possibility of interference cannot be excluded.

4 °C

Unopened Kits: Store at 4 °C for 6 months.
Microstrips Coated w/ Capture Antibody, 400x Streptavidin-HRP Wash Buffer (10x), Assay Diluent Ready-to-Use Substrate, Stop Solution: 6 Months at 4 °C.
Protein Standard, Biotinylated Detection Antibody: Lyophilized: 6 Months (if Reconstituted: 1 Month) at 4 °C.

6 months

Unopened Kits: Store at 4 °C for 6 months.
Microstrips Coated w/ Capture Antibody, 400x Streptavidin-HRP Wash Buffer (10x), Assay Diluent Ready-to-Use Substrate, Stop Solution: 6 Months at 4 °C.
Protein Standard, Biotinylated Detection Antibody: Lyophilized: 6 Months (if Reconstituted: 1 Month) at 4 °C.

For Research Use only

Vascular Endothelial Cell Growth Factor

MVCD1; VEGF; VPF; Vegf; Vegf120; Vegf164; Vegf188; Vpf; VEGF-A; VEGF164; eVEGF120; eVEGF164; vegf-a; vegf; vascular endothelial growth factor A; vascular endothelial growth factor A L homeolog; vascular endothelial growth factor; vascular endothelial growth factor precursor; VEGFA; Vegfa; vegfa.L; VEGF; vegf

Human VEGF, also known as Vascular Endothelial Growth Factor, is a 232 amino acid growth factor protein encoded from the VEGFA gene located at locus 6p12 on chromosome 6. After initial synthesis, the 26 residue signal sequence is cleaved from the N-terminal end, allowing for the 206 residue VEGF protein to fold properly into its mature form. VEGF is known to be a growth factor active in angiogenesis, vasculogenesis and endothelial cell growth. Through binding to the FLT1/VEGFR1 and KDR/VEGFR2 receptors, heparin sulfate and heparin, homodimeric VEGF is able to induce endothelial cell proliferation, promote cell migration, as well as inhibit apoptosis and induce permeabilization of blood vessels.

7422

P15692

RTK Signaling, Glycosaminoglycan Metabolic Process, Regulation of Cell Size, Tube Formation, Signaling Events mediated by VEGFR1 and VEGFR2, Platelet-derived growth Factor Receptor Signaling, VEGFR1 Specific Signals, VEGF Signaling

This particular immunoassay utilizes the quantitative technique of a Sandwich Enzyme-Linked Immunosorbent Assay (ELISA) where the target protein (antigen) is bound in a sandwich format by the primary capture antibodies coated to each well-bottom and the secondary detection antibodies added subsequently by the investigator. The capture antibodies coated to the bottom of each well are specific for a particular epitope on the Human VEGF cytokine while the user-added detection antibodies bind to epitopes on the captured target protein. Amid each step of the procedure, a series of wash steps must be performed to ensure the elimination of non-specific binding between proteins to other proteins or to the solid phase. After incubation and sandwiching of the target antigen, a peroxidase enzyme is conjugated to the constant heavy chain of the secondary antibody, allowing for a colorimetric reaction to ensue upon substrate addition. When the substrate TMB is added, the reaction catalyzed by peroxidase yields a blue color that is representative of the antigen concentration. Upon sufficient color development, the reaction can be terminated through addition of Stop Solution where the color of the solution will turn yellow. The absorbance of each well can then be read by a spectrophotometer, allowing for generation of a standard curve and subsequent determination of protein concentration.