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Femoral nerve block was performed using a linear ultrasound probe (HFL50 15–6 MHz; SonoSite Inc., Bothell, WA, USA) and a 5-cm, 18-gauge Contiplex Tuohy needle (B-Braun Ltd., Tochigi, Japan) [2].
We performed a study to determine, using ultrasonography, the actual location of the LA when TAP block was performed using landmark-based technique.
Bilateral transversus abdominis plane block was performed using 30 ml of 0.375% ropivacaine before surgery.
Thereafter, a paravertebral block was performed using lidocaine (Xylocaine, AstraZeneca, Brussels, Belgium), and a full thickness third-degree burn injury was inflicted on the flanks (n = 12) (15 20 % body surface area; painless by itself as cutaneous nerves for sensation of pain are destroyed).
In the PCSNB group, psoas compartment block was performed using modified Winnie technique [ 12].
An avidin biotin block was performed using reagents from Vector (Blocking kit, Cat. No. SP-2002, PeterboroUKh, UK).
Similar(52)
Blocking was performed using 10% normal donkey serum (NDS) in 1× PBS.
Endogenous peroxidases were deleted by incubation with 0.9% hydrogen peroxide for 15 min at room temperature (RT) and blocking was performed using 3% BSA for 1 h at RT. Antibodies were then applied in an optimal concentration overnight in a wet chamber.
Briefly, antigen retrieval was performed in 50 mM Tris buffer (pH 9.0) for 20 minutes, then blocking was performed using 10% normal goat serum diluted with PBS and 0.05% Tween-20 (PBS-T) for 1 hour at room temperature.
Cells were then fixed with 4% paraformaldehyde in PBS for 10 min at room temperature, washed with PBS and incubated with 0.1% NP-40 in PBS for 10 min. Blocking was performed using 10% goat serum (Vector Laboratories, Burlingame, CA) in PBS for 1 hr.
Blocking was performed using 1 % bovine serum albumin in PBS.
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