Table 1. (A) Compositions of E. coli Extraction Buffers, (B) Processing Conditions for the Eight 400 μg Aliquots of Standard Protein Mixture, (C) Filter-Aided Sample Preparation Buffer Components, and (D) E. coli Lysis Buffers Used to Evaluate eFASP with Surface Passivation.

(A) E. coli Extraction Buffers
extraction buffer	chaotrope	reductant/buffer
A	12 mM DCA, 12 mM N-lauroyl sarcosine	10 mM TCEP, 200 mM ABC, pH 8
B	12 mM DCA, 12 mM N-lauroyl sarcosine	10 mM DTT, 200 mM ABC, pH 8
C	4% SDS	10 mM DTT, 200 mM ABC, pH 8

(B) Standard Protein Mixture Processing Conditions
		buffer composition
aliquot	title	suspensiona	reductionb	alkylationc	quenchingd	digestione
1	DTT/IAN	ABC	DTT	IAN	DTT	ABC
2	DCA/DTT/IAN	DCA	DTT	IAN	DTT	DCA
3	DCA/DTT/4VP	DCA	DTT	4-VP	DTT	DCA
4	TCEP/IAN	ABC	TCEP	IAN	DTT	ABC
5	DCA/TCEP/IAN	DCA	TCEP	IAN	DTT	DCA
6	TCEP/4-VP	ABC	TCEP	4-VP	DTT	ABC
7	DCA/TCEP/4-VP	DCA	TCEP	4-VP	DTT	DCA
8	DCA/TCEP+DTT/4-VP	DCA	DTT/TCEP	4-VP	DTT	DCA

(C) Filter-Aided Sample Preparation Solubilization, Exchange and Digestion Buffers
	chaotrope/buffer	surfactant
solubilization buffer A	4% SDS, 200 mM ABC, pH 8, 7.5 mM TCEP	0.2% DCA
exchange buffer A	8 M urea, 100 mM ABC, pH 8	none
exchange buffer B	8 M urea, 100 mM Tris-HCl, pH 8	0.1% nOG
		0.1% DCA
		0.2% DCA
		0.1%DCA, 0.1% nOG
alkylation buffer A	8 M urea, 100 mM ABC, pH 8, 50 mM IAN
alkylation buffer B	100 mM ABC, pH 8, 30 mM IAN
FASP digestion buffer	50 mM ABC	none
eFASP digestion buffer	50 mM ABC	0.1%nOG
		0.1% DCA
		0.2% DCA
		0.1% DCA, 0.1% nOG
2D clean up digest buffer	100 mM ABC, pH 8	0.1% DCA

(D) Lysis Buffer Compositions for E. coli Evaluation of eFASP on Passivated Surfaces
lysis buffer	chaotrope	reductant/buffer
A	4% SDS, 0.2% nOG	7.5 mM TCEP, 200 mM ABC, pH 8
B	4% SDS, 0.1% DCA, 0.1% nOG	50 mM TCEP, 200 mM ABC, pH 8

^aABC = 25 mM ABC. DCA = 0.1% DCA, 25 mM ABC.

^bDTT = 22 mM DTT. TCEP = 5.5 mM TCEP. DTT/TCEP = 11 mM DTT/2.8 mM TCEP.

^cIAN = 30 mM IAN. 4-VP = 25 mM 4-VP.

^dDTT = 22 mM DTT.

^eABC = 25 mM ABC. DCA = 0.1% DCA, 25 mM ABC.