VWF (Human) ELISA Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Store the unopened kit at 2-8°C upon receipt and when it is not in use. -Keep microtiter plate in a sealed bag with desiccant to minimize exposure to damp air. -All reagents should be allowed to reach room temperature (18-25°C) before use.	-DO NOT DILUTE THE STANDARDS. -Rinse and flick the microtiter wells 5 times with deionized or distilled water. DO NOT USE TAP WATER. -It is recommended that the wells be read within 15 minutes following addition of Stop Solution.

Upstream tips
-Store the unopened kit at 2-8°C upon receipt and when it is not in use. -Keep microtiter plate in a sealed bag with desiccant to minimize exposure to damp air. -All reagents should be allowed to reach room temperature (18-25°C) before use.
Protocol tips
-DO NOT DILUTE THE STANDARDS. -Rinse and flick the microtiter wells 5 times with deionized or distilled water. DO NOT USE TAP WATER. -It is recommended that the wells be read within 15 minutes following addition of Stop Solution.

Publication protocol

LV halves (obtained as above; n = 5–9 per group) were either stimulated with mouse thrombin (100 nM, HaemTech; 30 min in HBS buffer, 37°C) to promote maximal vWF secretion, or left unstimulated (vehicle control). Two technical replicates of 50 μl each were removed from each well and a sandwich ELISA carried out as per manufacturer's instructions (Abnova, vWF ELISA kit, Taipei, Taiwan) to quantitate vWF extruded from the LV endocardial surface. A standard curve with known vWF concentrations (2.5–80 mU) was run in parallel. Data from the technical replicate wells were averaged to obtain a mean value per mouse left ventricle. To measure endocardial vWF protein levels (n = 4 per group), the LV was placed endocardial surface up onto a modified 384-well plate and Lysis M buffer applied (75 mL, 30 min); lysate was centrifuged then 200 μg protein per sample was processed for vWF ELISA as above.

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Manufacturer protocol

Download the product protocol from Abnova for VWF (Human) ELISA Kit below.

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