Human Peroxisomal testis-specific protein 1 (PXT1) ELISA Kit

Genes reported to be crucial for spermatogenesis are often exclusively expressed in the testis. The putative amino acid sequence encoded by the cDNA of the Pxt1 gene contains a conserved Asn-His-Leu (NHL) -motif at its C-terminal end, which is characteristic for peroxisomal proteins. Pxt1-EGFP fusion protein is co-localized with known peroxisomal marker proteins in transfected NIH3T3 cells. The peroxisomal targeting signal NHL is functional and responsible for the correct subcellular localization of the Pxt1-EGFP fusion protein. In male germ cells peroxisomes were reported only in spermatogonia. The Pxt1 gene is so far the first gene coding for a putative peroxisomal protein which is expressed in later steps of spermatogenesis, namely in pachytene spermatocytes.

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Human Peroxisomal testis-specific protein 1 (PXT1) ELISA Kit has high sensitivity and excellent specificity for detection of Human PXT1. No significant cross-reactivity or interference between Human PXT1 and analogues was observed.

This Human Peroxisomal testis-specific protein 1 (PXT1) ELISA Kit employs a two-site sandwich ELISA to quantitate PXT1 in samples. An antibody specific for PXT1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyPXT1 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for PXT1 is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of PXT1 bound in the initial step. The color development is stopped and the intensity of the color is measured.

• Do not mix components from different kit lots or use reagents beyond the kit expiration date.• Allow all reagents to warm to room temperature for at least 30 minutes before opening.• Pre-rinse the pipet tip with reagent, use fresh pipet tips for each sample, standard and reagent to avoid contamination.• Unused wells must be kept desiccated at 4 °C in the sealed bag provided.• Mix Thoroughly is very important for the result. It is recommended using low frequency oscillator or slight hand shaking every 10 minutes.• It is recommended that all samples and standards be assayed in duplicate or triplicate.