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At the same time blocking studies were performed.
In group B (n = 4), non-radioactive TAF (3.1 nmol, in PBS) was injected 5 min prior to the injection of [18F]TAF (17.8 ± 1.7 MBq, 1.2 ± 0.1 nmol, in 10% EtOH in PBS); otherwise, the experiments were performed as in group A. Two blocking studies were performed in order to ensure the specificity of the binding at the tumour site.
To avoid uptake, antibody blocking studies were performed at 4°C (30 min).
To verify the specificity of PI3K-C2 immunoreactivity, blocking studies were performed by pre-incubation of each antibody with the protein fragment used to raise it prior to its application in IHC.
To this end, blocking studies were performed in arthritic rats by pre-administration of 5 mg/kg unlabelled PK11195 (Sigma Aldrich Chemie BV) five minutes prior to the injection of [C]DPA-713 or [F]DPA-714.
To verify the specificity of the CTCF immunoreactivity, blocking studies were performed by preincubation of the primary antibody (at different dilutions) for 2 h at room temperature with the protein fragment used to raise it (Santa Cruz Biotechnology, sc-5916 P) prior to its application to the tissues according to the supplier data sheet protocol.
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Receptor-blocking studies were performed using 100 μg of unlabelled BN(1 14) co-injected with the corresponding radiolabelled BN analogue.
Using IgG concentrations within the range of the IC50's for R217, R218 and R256 (Table 1), and [35S]-metabolically labeled native EBA-175, blocking of binding studies were performed such that the proportion of mAb R217 equaled one minus the proportion of mAb R218 or R256.
If a neuromuscular transmission defect was suspected, as in case of recent use of neuromuscular blocking agents, low frequency (3Hz) repetitive stimulation studies were performed with stimulation of the right ulnar nerve and recording from abductor digiti minimi and stimulation at wrist.
Antigen adsorption studies were performed using COX-2 blocking peptide.
First, the fMRI studies were performed with block design paradigm in this work, and temporal redundancy from the spatial-temporal frequency domain may have been exploited better for such data type.
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