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  • Quick histochemical staining method for measuring lactate dehydrogenase C4 activity in human spermatozoa.

Quick histochemical staining method for measuring lactate dehydrogenase C4 activity in human spermatozoa.

Acta histochemica (2015-03-22)
Zhaolei Cui, Liangyuan Chen, Yaohua Liu, Zhangxin Zeng, Fenghua Lan
ABSTRACT

The enzyme activity of lactate dehydrogenase C4 (LDH-C4, due to tetrameric nature of C-subunit) has been proposed as an important parameter in evaluating sperm motility and semen quality. A novel histochemical staining method for detecting LDH-C4 activity in human spermatozoa is described in this report. The staining working solution comprises sodium 2-hydroxybutyrate (an affinity substrate of LDH-C4), nitrotetrazolium blue chloride (NBT), nicotinamide adenine dinucleotide (NAD) and naphthol blue. The positive products were purple black lumps concentrated in the neck segment of the spermatozoa and weakly in the middle piece. A normal reference range for the integral enzyme activity was constructed from 120 healthy males based upon the scoring criteria. The study further compared the staining method with the routine spectrophotometry technique in terms of the results of 96 cases with infertile status. Moreover, we found the down-regulated LDH-C4 expression was significantly correlated with the lowered enzyme activity (r=0.865, P=0.000). Our data suggest that the histochemical staining method hallmarks a relatively high accuracy and may be a better alternative for measuring LDH-C4 activity in human spermatozoa.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
1-Naphthol, puriss. p.a., reag. Ph. Eur., ≥99% (GC)
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1-Naphthol, BioXtra, ≥99%
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1-Naphthol, ReagentPlus®, ≥99%
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DAPI, for nucleic acid staining
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β-Nicotinamide adenine dinucleotide hydrate, ≥98%, BioUltra, from yeast
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β-Nicotinamide adenine dinucleotide hydrate, Grade AA-1
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β-Nicotinamide adenine dinucleotide hydrate, ≥99%
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β-Nicotinamide adenine dinucleotide hydrate, purified by column chromatography, ≥99%
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β-Nicotinamide adenine dinucleotide hydrate, suitable for cell culture, ≥96.5% (HPLC), ≥96.5% (spectrophotometric assay), from yeast
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β-Nicotinamide adenine dinucleotide hydrate, ≥96.5% (HPLC), ≥96.5% (spectrophotometric assay), from yeast
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β-Nicotinamide adenine dinucleotide hydrate, ≥95% (HPLC)
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β-Nicotinamide adenine dinucleotide, pkg of 20 mg (per vial)
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