Table 2. Amount of antibody penetrance obtained with various detergents and incubation times.

exp	perfuse	detergent	thick-ness	1° Ab time	2° Ab time	penetrance	quality of labeling
initial	yes	0.5% Tx	400 μm	7d	7d	fair	–
1	no	1% Tx	300 μm	3.5d	2d	fair	–
1	yes	1% Tx	300 μm	3.5d	2d	fair	–
1	no	1% NP40	400 μm	3.5d	2d	fair	–
1	yes	1% NP40	400 μm	3.5d	2d	fair	–
2	no	0.5% Chp	350 μm	5d	4.5d	poor	–
2	no	1% Chp	350 μm	5d	4.5d	fair	–
2	no	1% SDS in pre-tr, then 2% Tx	350 μm	5d	4.5d	fair	poor morphology
2	no	2% Tx	300 μm	5d	4.5d	fair	–
3	no	0.5% Tw	350 μm	10d	10d	fair	–
3	no	0.5% Tx	350 μm	10d	10d	complete	ok
3	no	1% Tx	350 μm	10d	10d	complete	good
3	no	2% Tx	350 μm	10d	3d	complete	ok
3	no	2% Tx	350 μm	10d	7d	complete	good
3	no	2% Tx	350 μm	10d	10d	complete	best

Conditions for each experiment (exp) are listed below.

initial: permeabilize 2 hrs, RT; incubate in 250 μl in 96-well plate

exp 1: permeabilize 2 hrs, 37°C; incubate in 250 μl in 96-well plate

exp 2: permeabilize 5 hrs, 37°C; incubate in 250 μl in 96-well plate

exp 3: permeabilize 4.5 hrs, RT; incubate in 400 μl in small centrifuge tubes

Ab, antibody; tr, treatment; Tx, Triton X-100; Tw, Tween-20; Chp, CHAPS