Mouse Fibronectin PicoKine™ ELISA Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Store at 4°C for 6 months, at -20°C for 12 months. -Avoid multiple freeze-thaw cycles. -The standard solutions are best used within 2 hours.	-The diluted Avidin-Biotin-Peroxidase Complex (ABC) solution should not be prepared more than 1 hour prior to the experiment. -Prepare the substrate solution immediately before use or bring the pre-made substrate to room temperature. -Sodium azide is an inhibitor of HRP. Do not include the azide in buffers or wash solutions if HRP-labeled conjugate is used for detection.

Upstream tips
-Store at 4°C for 6 months, at -20°C for 12 months. -Avoid multiple freeze-thaw cycles. -The standard solutions are best used within 2 hours.
Protocol tips
-The diluted Avidin-Biotin-Peroxidase Complex (ABC) solution should not be prepared more than 1 hour prior to the experiment. -Prepare the substrate solution immediately before use or bring the pre-made substrate to room temperature. -Sodium azide is an inhibitor of HRP. Do not include the azide in buffers or wash solutions if HRP-labeled conjugate is used for detection.

Publication protocol

Plasma levels of cytochrome C and HMGB1, S100A, fibronectin, and hyaluronan were assayed by ELISA according to the manufacturer’s recommendations. The ELISA kits for cytochrome C, HMGB1 and S100A were from www.antibodies-online.com. The kit for fibronectin was from Boster (Pleasanton, CA) and the kit for hyaluronan was from R&D Systems (Minneapolis, MN). Total double stranded DNA was isolated from plasma using a QIAamp DNA blood minikit (Qiagen, Valencia CA) following the manufacturer’s recommendations. DNA in the plasma was determined spectrophotometrically by measuring the absorbance of the sample at 260nm (A260) and adjusting the measurement for turbidity (measured by absorbance at 320nm). The concentration of DNA (µg/mL) = (A260 value – A320value) × the dilution factor × 50µg/mL (i.e., A260 of 1.0 = 50µg/mL double stranded DNA). The total yield of DNA was obtained by multiplying the DNA concentration by the sample volume.

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Manufacturer protocol

Download the product protocol from BosterBio for Mouse Fibronectin PicoKine™ ELISA Kit below.

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