Protocol for in vivo bioorthogonal click chemistry by Ac4MaNAz

[Nilesh Meghani]

Hello All,

I am working on bioorthogonal click chemistry. So, if anyone can please help with the protocol for the in vivo labelling of Ac4MaNAz. ? Thank you for your time.

Kind Regards,

Nilesh

[Stephen Roper]

Hi Nilesh,

  I do In vitro labeling with Ac4MaNAz for downstream MS analyses- perhaps I can help. What type of assay will you be doing post-labeling? Live-cell imaging? It is my understanding that for In vivo labeling you will need a copper-free system, such as those available in commerical kits (http://tools.invitrogen.com/content/sfs/manuals/mp10405.pdf). Obviously you need to purchase the AC4MaNAz as well as an alkyne-based substrate for detection in addition to this copper-free reagent (DIBO). The Invitrogen website has been a great help to me in developing click chemistry protocols (http://www.invitrogen.com/site/us/en/home/References/Molecular-Probes-The-Handbook/Reagents-for-Modifying-Groups-Other-Than-Thiols-or-Amines/Click-Chemistry.html).  Hope this helps, SR. 

[Stephen Roper]

After further review, I believe I was mistaken in my first post. In copper-free click chemistry the detection reagent is NOT an alkyne, it is the DIBO reagent conjugated to a fluorphore/hapten (as listed in table 2 in that pdf I posted a link to before). So basically you would "feed" your cells the AC4ManNAz, then treat with the DIBO-conjugated fluuorphore/hapten for detection (i.e. no alkyne-based substrate for detection). -SR

On Tuesday, August 6, 2013 1:57:41 AM UTC-4, Nilesh Meghani wrote:

[Nilesh Meghani]

Dear Dr. Roper,

I understood that. Thank you for your time.

Kind Regards,

Nilesh