For the preparation of nuclear extracts, frozen liver pieces were homogenized in homogenization buffer (20 mM HEPES, 70 mM sucrose, 220 mM Mannitol, 2 mM EDTA, 0.5 mg/ml BSA, 0.1 mM PMSF, 1.0 mM DTT, pH 7.5)
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8 Where results are unclear, serum MMA and homocysteine levels (both of which accumulate with B12 deficiency as discussed above) can also be used to confirm a diagnosis, although homocysteine is not specific to B12 deficiency, as it may also be caused by deficiencies in other B vitamins