Lysis Buffers Comprising Cyanate or Thiocyanate and a Detergent
The present disclosure provides cell lysis buffers and method of use. Cell lysis buffers may comprise an aqueous solution comprising a buffering agent, a detergent, and cyanate or thiocyanate. The cell lysis buffers may be free or substantially free of urea and/or carbonic acid. Such lysis buffers may be used to lyse cells, e.g., in tissues. The cell lysis buffers may be suitable for use with isotachophoresis.
1 . A composition comprising:
a) a buffer in aqueous solution;
b) a detergent; and
c) about 5 mM to about 100 mM cyanate or thiocyanate;
wherein a relative concentration of the cyanate or the thiocyanate to urea in the composition is at least about 1:800, at least about 1:600, at least about 1:400, at least about 1:200, at least about 1:10, at least about 1:1, at least about 100:1, at least about 10,000:1, or at least about 1,000,000:1.
2 . The composition of claim 1 , wherein the detergent is an ionic detergent.
3 . The composition of claim 2 , wherein the ionic detergent is a cationic, anionic, zwitterionic, or amphoteric detergent.
4 . The composition of claim 1 , wherein the detergent is a surfactant.
5 . The composition of claim 1 , wherein the detergent is a non-ionic detergent.
6 . (canceled)
7 . The composition of claim 1 , comprising no more than about 500 mM urea, no more than about 10 mM urea, no more than about 2 mm urea, no more than about 100 nM urea, or no more than about 1 nanomolar urea.
8 . The composition of claim 1 , wherein the composition is free or essentially free of urea.
9 . The composition of claim 1 , wherein the composition is free or essentially free of guanidinium.
10 . The composition of claim 1 , further comprising about mM to 25 mM cyanate or thiocyanate, about 25 mM to 50 mM cyanate or thiocyanate, or about 50 mM to 100 mM cyanate or thiocyanate.
11 . The composition of claim 1 , wherein the cyanate or the thiocyanate comprises sodium cyanate, sodium thiocyanate, potassium cyanate, or potassium thiocyanate.
12 . The composition of claim 1 , further comprising an ionic salt.
13 - 14 . (canceled)
15 . The composition of claim 12 , wherein the ionic salt is present at about 10 mM to 40 mM.
16 . The composition of claim 1 , wherein the buffer comprises phosphate, Tris, Bis-Tris, e-aminocaproic acid, or HEPES.
17 - 19 . (canceled)
20 . The composition of claim 1 , wherein the aqueous solution has a pH between about 3.5 and about 9.0.
21 . (canceled)
22 . The composition of claim 1 , wherein the buffer has a concentration between about 10 mM and about 200 mM.
23 . (canceled)
24 . The composition of claim 1 , wherein the detergent comprises a dodecyl sulfate or ethyl trimethylammonium bromide (“ETMAB”).
25 . The composition of claim 24 , wherein the dodecyl sulfate comprises sodium dodecyl sulfate.
26 . The composition of claim 1 , further comprising a reducing agent.
27 . The composition of claim 26 , wherein the reducing agent comprises DTT (dithiothreitol), DTE (dithioerythritol), L-glutathione (GSH), TCEP (Tris (2-Carboxyethyl) phosphine hydrochloride), or beta-mercaptoethanol.
28 . The composition of claim 1 , further comprising a protease.
29 - 99 . (canceled)