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Next, an IF-IHC procedure is carried out as follows: after all IF-IHC reagents are pre-cooled to 4°C, blocking solution (comprised of 1% normal goat serum "NGS," Sigma, and 1% normal swine serum "NSS," Invitrogen, Grand Island, NY, added to Common Antibody Diluent, BioGenex) is added to the sections for 5 minutes.
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The membranes were incubated in blocking solution comprising 5% nonfat milk in TBST at room temperature for 1.5 h and then incubated with the primary antibodies diluted in blocking solution individually.
Completely aspirate off the blocking solution and add 30 μl of primary antibody solution per well.
Following removal of blocking solution, primary antibodies in wash solution were incubated with the tissue slices.
Peroxidase blocking solution (DAKO S 2023) was used for blocking of endogenous peroxidises.
The dilution of antibodies was performed in blocking solution.
(1 100 in blocking solution).
Discard the blocking solution.
diluted 1 100 in blocking solution.
A solution comprising of six factors was chosen.
As shown by Figure 1, there were three haplotype blocks generated including Block 1 comprised of SNP 1 4, Block 2 comprised of SNP 5 6 and Block 3 comprised of SNP 8 9.
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