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The rule is to move at least one inch from the previous injection in the same zone and to wait at least seven to fourteen days before using any specific point again

After the membranes were incubated in blocking buffer at room temperature for 2 h, they were washed 3 times in tris-buffered saline and tween 20 (TBST) solution, 5 min each, and then incubated with primary antibodies overnight at 4 C

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All immune-related findings should be interpreted as supportive rather than conclusive

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