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This increase in binding was prevented by inactivation of HMGA1a in the nuclear extracts using an antibody against it (Figures. 2A and 2B).
For immunohistochemistry, paraffin sections were reacted with rabbit anti-periostin antibody or anti-Ki67 antibody (YLEM, Rome, Italy) or anti-fibronectin antibody (Sigma-Aldrich, St Louis, MO) after the endogenous peroxidase had been blocked with 3% H2O2, and nonspecific binding was prevented by treatment with 2% bovine serum albumin (BSA; Nacalai Tesque, Kyoto, Japan) in Tris-buffered saline.
Non-specific binding was prevented by incubation with pure goat serum for 30 min on ice.
Non-specific binding was prevented by incubation of the slides with 30% normal rabbit serum in PBS during 30 min at room temperature.
Endogenous peroxide activity was inhibited by incubation with 3% H202, and nonspecific antibody binding was prevented by treating with 10% normal goat serum and avidin/biotin blocking solutions.
Endogenous peroxidase activity was blocked with 3% hydrogen peroxide and nonspecific binding was prevented by incubation with 3% normal serum for 1 h at room temperature.
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In its normal inactive state, RON is bound to Hyal2, and when Hyal2 binding is prevented by JSRV Env, RON becomes constitutively active in the absence of MSP stimulation.
Several VCP adaptors and coadaptors whose binding is prevented by VCP phosphorylation (Li et al., 2008; Madsen et al., 2008; Zhao et al., 2007) have, however, been identified.
In the natural chromosome configuration, E. coli cells lacking these three helicases are viable only if the stability of the RNA polymerase is compromised and RecA binding is prevented by an RecF mutation.
Enterocyte damage, expressed as circulating levels of ileal lipid binding protein, was prevented by high-lipid feeding compared with animals that received a low-lipid composition or were fasted (3.7 ± 0.3 vs 4.9 ± 0.5 vs 8.0 ± 1.1 pg/ml; P < 0.05 respective P < 0.0001).
Nonspecific binding of proteins was prevented by treating the membranes with 5% nonfat dry milk in Tris-buffered saline/0.1% Tween 20 for 2 hrs at room temperature.
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