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To adjust for variation in the volume of phosphate buffered saline (PBS) used during the CVL (10 or 12 ml) concentrations determined from samples with lower dilution (10 ml) were divided by a factor of 1.2.
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Rabbits were anesthetized with medetomidine (Domitor 0,3 mg/kg, Orion, Finland) and ketamine (Ketalar 20 mg/kg, Pfizer, Finland) and the total dose of 1×1011 virus particles in 1 ml was divided to ten 100 µl injections.
The resulting supernatant (∼ 4 ml) was divided into aliquots for storage at −80°C.
The eluate between Vo (15 ml) and Vt (105 ml) was divided into six pools (15 ml each), freeze-dried and reconstituted in a small volume of TNC buffer.
Briefly, the cells in 1 ml PBS were divided equally into two portions, one for total sterol (with saponification) and the other for free sterol (without saponification).
Both SAHA and ML-60218 were divided into aliquots and stored at –20°C.
To generate whole cell lysates and soluble fractions, 10-mL cultures were divided and pelleted by centrifugation for 15 min at 4°C and 3500 rpm.
The pooled and dialyzed S-Sepharose Elba fractions (3 ml first experiment or 6 ml second experiment) were divided into two equal parts and incubated in ∼1 ml aliquots with 3× Elba-site or 3× Elba-M3-site (50 μl first or 80 μl second) affinity beads.
20 ml crude extracts from 100 ml culture broth were divided into four equal parts.
Best results could be observed when 77 mL of hexane were divided in four consecutive extraction steps with 18 min each.
CFA (0.08 ml -inflamed rats were divided into four groups (n = 7 per group): RU486 (Sigml -inflamedA, vehicle + sham EA, rats6 + EA and vehicle + EA.
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