Human Osteopontin ELISA Kit (ab100618)

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

The serum and joint fluid samples were subjected to ELISA to determine the concentration of OPN, performed according to the manufacturer's protocol of the OPN ELISA kit (cat. no. ab100618; Abcam, Cambridge, UK). Briefly, 50 µl of the standard samples at 0, 74.07, 222.2, 666.7, 2,000, 6,000 and 18,000 pg/ml were added into different wells on the ELISA plate. Next, 10 µl sample and 40 µl dilution solution was added into each well. With the exception of the blank well, each well was treated with 100 µl biotinylated antibodies and incubated at 37°C for 1 h. Subsequent to washing five times, 50 µl substrate A and 50 µl substrate B were added into each well, incubated at 37°C for 15 min, and followed by addition of 50 µl stop solution in order to terminate the reaction. Optical density values were measured at a wavelength of 450 nm within 15 min.

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Manufacturer protocol

Download the product protocol from Abcam for Human Osteopontin ELISA Kit (ab100618) below.

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