Human Serpin E1/PAI-1 Quantikine ELISA Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Store the unopened kit at 2-8 °C. -Bring all reagents to room temperature before use. -Reconstitute the Human Cytochrome c Standard with deionized or distilled water. Allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making dilutions.	-When mixing or reconstituting protein solutions, always avoid foaming. -Substrate Solution should remain colorless until added to the plate. -Keep Substrate solution protected from light. -Stop Solution should be added to the plate in the same order as the Substrate Solution. -Wells that are green in color indicate that the Stop Solution has not mixed thoroughly with the Substrate Solution.

Upstream tips
-Store the unopened kit at 2-8 °C. -Bring all reagents to room temperature before use. -Reconstitute the Human Cytochrome c Standard with deionized or distilled water. Allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making dilutions.
Protocol tips
-When mixing or reconstituting protein solutions, always avoid foaming. -Substrate Solution should remain colorless until added to the plate. -Keep Substrate solution protected from light. -Stop Solution should be added to the plate in the same order as the Substrate Solution. -Wells that are green in color indicate that the Stop Solution has not mixed thoroughly with the Substrate Solution.

Publication protocol

Double antibody sandwich enzyme-linked immunosorbent assay (ELISA) was used to measure serum protein concentrations of tPA, PAI-1, BDNF, proBDNF, TrkB and p75NTR. The human tPA, PAI-1 and BDNF levels were measured by ELISA kits [tPA (DTPA00; R&D Systems; USA); PAI-1 (DSE100; R&D Systems; USA); BDNF (DBD00, R&D Systems; USA)] and proBDNF, TrkB and p75NTR levels were assessed using DuoSet human ELISA Development System (proBDNF: DY3175, TrkB: DYC397-2, p75NTR: DY367 and Ancillary Reagent: DY008; R&D Systems; USA). Protocols were performed according to the manufacturer’s instructions. In order to minimize assay variance, each protein concentrations of all subjects were measured on the same day. All experiments were performed in duplicate. The sensitivity of tPA, PAI-1 and BDNF assay were 1.40-16.1 pg/mL, 0.014-0.142 ng/ml and less than 20 pg/mL, respectively; while the sensitivity of proBDNF, TrkB and p75NTR assay were not given exactly in the specification. No significant cross-reactivity or interference was observed in each assay.

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Manufacturer protocol

Download the product protocol from R&D Systems for Human Serpin E1/PAI-1 Quantikine ELISA Kit below.

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