Mouse RANKL ELISA Kit (TNFSF11) (ab100749)

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

BMMSCs were plated at 5 × 104 cells/well in 24-well culture plates in α-MEM and were then treated with RANKL (30 ng/mL) and M-CSF (10 ng/mL) for 5 days. The media were changed every 48 hrs. After 5 days of culture, cells were xed with 4% paraformaldehyde (PFA) and stained with tartrate-resistant acid phosphatase (TRAP; Sigma). TRAP-positive cells were imaged under a phase-contrast microscope and counted. Similarly, osteoclast activity was determined by measuring TRAP5b activity in plasma using an ELISA kit from MyBioSource as per the manufacturer’s instructions.

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Manufacturer protocol

Download the product protocol from Abcam for Mouse RANKL ELISA Kit (TNFSF11) (ab100749) below.

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