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Endogenous avidin and biotin were blocked using the Vector Avidin/Biotin Blocking Kit (Vector Laboratories, Inc., Burlingame, CA), and further blocking was achieved using swine serum.
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Paracervical block was achieved using a 27-gauge spinal needle.
A stratified (on center and BMI), blocked randomization was achieved using a random number program in S-plus written by the study biostatistician (R F B). Numbered, sealed envelopes were opened by the Study Coordinator at the time of randomization.
To investigate the importance of OX40 signals for physiological CD4+ T-cell responses, an endogenous antigen-specific population of CD4+ T cells that recognise the 2W1S peptide was assessed and temporal control of OX40 signals was achieved using blocking or agonistic antibodies (Abs) in vivo.
Blocking of HIF-1α was achieved using the commercially available inhibitors CTM (Chetomin, NSC 289491, Invitrogen), and digoxin (Sigma-Aldrich, Buchs, Switzerland).
Blocking of the membrane was achieved using 5% nonfat milk in TTBS (pH 7.6).
Blocking of endogenous peroxidase activity was achieved using 3% hydrogen peroxide for 10 min, and then sections were blocked using 5% albumin from bovine serum (BSA; Sigma-Aldrich) for 30 min.
Blocking of leptin's expression was achieved using siRNA against leptin and transfection with liposomes.
In addition, subjects were required to have an Adult Myopathy Assessment Tool (AMAT) 5 score from 14 to 41. Subject group randomization was achieved using stratified block randomization based on AMAT scores with a block size of four.
The experiment required reproducible longitudinal alignment of the bone during compression testing, and this was achieved using a mounting block.
Staining was achieved using the Vectastain Elite ABC kit (universal), containing blocking serum, biotinylated secondary antibody and ABC reagent (Vector Laboratories, Peterborough, UK).
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