Human PlGF Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human PlGF Immunoassay is a 3.5 or 4.5 hour solid phase ELISA designed to measure human PlGF in cell culture supernates, serum, plasma, and urine. The immunoassay kit contains E. coli-expressed recombinant human PlGF and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate recombinant human PlGF accurately. Results obtained using natural human PlGF showed dose curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural human PlGF.

Preparation and Storage

Background: PlGF

PlGF (placenta growth factor) is a homodimeric growth factor that competes with VEGF for binding to VEGF R1/Flt-1. It therefore increases the availability of VEGF to bind to VEGF R2/KDR/Flk-1 and trigger angiogenesis. It can form heterodimers with some forms of VEGF and decrease the angiogenic effect of VEGF on VEGF R2. Circulating PlGF levels increase during pregnancy, reaching a peak in mid-gestation; this increase is attenuated in preeclampsia. PlGF induces monocyte activation and migration as well as production of inflammatory cytokines and VEGF. These activities facilitate wound, bone fracture, and cardiac repair, but also contribute to inflammation in active sickle cell disease and atherosclerosis. PlGF can also inhibit TIMP3 expression in the spleen, leading to immune triggering of hypertension.

Assay Procedure

Refer to the product datasheet for the complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.

1.     Prepare all reagents, standard dilutions, and samples as directed in the product insert.

2.     Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

3.     Add 50 μL of Assay Diluent to each well.

4.     Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.

5.     Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

6.     Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.

7.     Aspirate and wash 5 times.

8.     Add 100 μL Substrate Solution to each well.

9.     Add 100 μL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.