Rat PAI1 ELISA Kit (SERPINE1) (ab198509)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Equilibrate all reagents to room temperature (18-25°C) prior to use.	-Avoid foaming or bubbles when mixing or reconstituting components. -Ensure plates are properly sealed or covered during incubation steps. -Complete removal of all solutions and buffers during wash steps is necessary to minimize background.	-Unused well strips should be returned to the plate packet and stored at 4°C.

Upstream tips
-Equilibrate all reagents to room temperature (18-25°C) prior to use.
Protocol tips
-Avoid foaming or bubbles when mixing or reconstituting components. -Ensure plates are properly sealed or covered during incubation steps. -Complete removal of all solutions and buffers during wash steps is necessary to minimize background.
Downstream tips
-Unused well strips should be returned to the plate packet and stored at 4°C.

Publication protocol

After the CUMS procedures and behavioural tests were completed, rats from the above four groups were anesthetized with 10% chloral hydrate. We used a stereotaxic apparatus with ear bars to hold the anesthetized rats in place. We made a longitudinal incision (approximately 2 cm) beginning between the ears and ending caudally. We then directly inserted a microliter syringe needle (Gaoge, China) from the lateral posterior of the foramen magnum into the cisterna magna and withdrew CSF. We centrifuged the CSF at 2000 rpm at 4 °C for 20 min and stored it at −80 °C until use for further assay. After withdrawing the CSF, we quickly opened the rat, inserted a disposable venous blood lancet directly into the left ventricle of heart, and collected the blood into a plain tube. The blood was centrifuged at 2000 rpm at 4 °C for 20 min, and the supernatants (serum) were frozen at −80 °C. The active PAI-1 concentrations in the serum and CSF were measured using enzyme linked immune sorbent assay [ELISA, PAI-1 (SERPINE1): ab198509, abcam] according to the manufacturer’s instructions. Each sample was tested in duplicate. The minimum detectable dose was 0.08 ng/ml.

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Manufacturer protocol

Download the product protocol from Abcam for Rat PAI1 ELISA Kit (SERPINE1) (ab198509) below.

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