Computational protocol: NIMA-related kinase 1 (NEK1) regulates meiosis I spindle assembly by altering the balance between α-Adducin and Myosin X

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Protocol publication

[…] Whole testis protein and oocyte protein were extracted by sonication in RIPA buffer. Samples were boiled for 5 min in sample buffer, electrophoresed on SDS-polyacrylamide gels (8%) and transferred to nitrocellulose membranes. Primary antibody incubation was performed overnight at 4°C at 1:1000 dilution (antibodies are the same used in spindle staining). Incubation with secondary antibodies was performed for two hours at room temperature (secondary HRP conjugated antibodies were obtained from Pierce, Life Technologies). Signal-detection was carried out using the superSignal substrate (Thermo Scientific). Loading control was performed using GAPDH-HPR (PA1-987-HRP, from ThermoFisher). Images were captured with BIO RAD Image Lab 5.1and analyzed by ImageJ version 1.49v ( […]

Pipeline specifications

Software tools Image Lab, ImageJ
Application Microscopic phenotype analysis
Organisms Homo sapiens, Mus musculus
Diseases Anemia, Carcinoma, Infertility, Polycystic Kidney Diseases, Muscular Diseases
Chemicals Tyrosine