We are TRAP staining mouse femurs, and eventhough the osteoclast staining looks specific they are staining yellow instead of red. Any thoughts?
In my case, it looked pinkish to red in mouse femur. Some cells stained red, otherwise pinkish. May be some pH issue, I am not sure. Use some different protocol.
What stain are you using? AS-TR red or something else?
Also I recommend not staining background, as if your staining is not strong enough, it does not fade against background stain.
To increase stain intensity you can increase the reaction time. Also ensure your stains are fresh, so use them immediately after taking them out of freezer and making solutions.
What test are you using? Kit from Sigma? And How are your Osteoclasts looking alike. Large cells with more than one nuclei? Be sure to use the right fixation solution! Here the pH is important. And do not rinse with PBS after fixation! The PBS is crossreating with the staining solution. AFTER fixation wash with water. To solve your problem some more details are necessary!
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