I have a series of thiol-modified aptamers in their oxidized form and want to reduce them using Tris [2-carboxyethyl] phosphine aka TCEP. As far as I have noticed, this chemical is sold in its hydrochloride salt form.
I was wondering if using the hydrochloride salt of the TCEP instead of TCEP itself would cause a problem due to the very acid pH caused by hydrochloride salt.
Anyone have any thoughts/suggestions?
TCEP (Tris(2-carboxyethyl) phosphine) is a reducing agent commonly used in biochemistry and molecular biology to reduce disulfide bonds in proteins and peptides. It is also used to reduce thiol-modified oligonucleotides, such as thiolated DNA or RNA oligos.
TCEP exists as a free thiol-reactive compound, but it can also be converted to a hydrochloride (HCl) salt form, known as TCEP+ HCl, for ease of handling and storage. The TCEP+ HCl salt is water-soluble and more stable than the free TCEP in solution.
When using TCEP or TCEP+ HCl for reducing thiol-modified oligos, the steps typically involve:
It is important to use appropriate protective measures, such as wearing gloves and working in a well-ventilated area when handling TCEP or its derivatives, as they are reactive compounds. Additionally, always follow the specific protocols and guidelines provided by the manufacturer or established scientific literature for the reduction of thiol-modified oligonucleotides.
Samsul Islam thank you for sharing the procedure.
My main concern is the acidic nature of the final solution that may affect the metal surface I will use for immobilizing my oligos.
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