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In addition to my previous email, I think that for the following experiments that I'm planning to conduct, I'd prefer to use an easylink conjugation kit for directly labelling the mouse monoclonal antibody (anti-geminin ab104306) rather than involving another secondary antibody into the system (I've got enough from them already!) =)
This primary antibody (ab104306) has a concentration of 0.36mg/mL, and I'm wondering whether is feasible or not to conjugate this antibody to TRITC (ab102915). The issue here is that it's a little bit more diluted than than the optimal concentration range suggested in the website (0.5mg-5 mg/mL), therefore I'd appreciate your suggestions about this matter.
Shall I use a mouse isotype control? if so, which one would be best provided that I'll be using this product ab104306?
Lookijng forward to hear from you.
Asked on Apr 10 2012
I'm sorry for the delay in getting back to you, we have been closed over the Easter holidays.
I have had a look at the secondary antibodies you have suggested as trying as an alternative to the one currently used (ab60314). All seem fine for the application you hope to use. If you would like I can send this out to you free of charge. Unless you have a preference for a different one, I'd send out ab96895.You mentioned that you were in Ireland? Would you still like for this replacement secondary antibody to be sent to the address I have for you:
I am not sure how the antibody concentration of anti-Geminin antibody ab104306 would affect the use in conjunction with the conjugation kit (ab102915). If will forward this query to the source of this kit and will let you know as soon as I have a response. As you will be conjugating directly, I would suggest that using an isotype control would be quite important asyou are not able to do a "no primary" control experiment to ascertain the non-specificity observed.Mouse monoclonalab81032 should be suitableand would becompatiblewiththe conjugationkit.
Can I ask, how are you going to detect the S phase (with Alexa Fluor 647), are you using another antibody? In which case, what antibodies do you intend to use?
How have your subsequent experimentsprogressed? Have you used the HeLa cellswith adherent slides?
I hope this information has been of help.I look forward toreceivingyour response.
Answered on Apr 10 2012