mouse monoclonal cmyc  (Novus Biologicals)

Journal: Journal of medical genetics

Article Title: De novo variants in SLC12A6 cause sporadic early-onset progressive sensorimotor neuropathy

doi: 10.1136/jmedgenet-2019-106273

Figure Lengend Snippet: (A) Pedigrees of the three unrelated affected patients (closed symbols) with de novo variants in SLC12A6 and healthy family members (open symbols). Wildtype is depicted as wt. (B) Images of patient 2 at 14 years show muscle atrophy predominantly in distal lower limbs, contractures of fourth and fifth fingers and flat feet. (C) Conservation of KCC3 Arg207 and Tyr679 among the different members of the SLC12A (cation-chloride) cotransporters. The arginine and tyrosine residues are boxed. (D) Conservations of same residues within KCC3 transporters from different species. (E) K+ influx measured in oocytes injected with water, KCC3-WT, Arg270His, Arg270Cys and Tyr679Cys. Oocytes were incubated under isotonic (grey bars) or hypertonic (black bars) conditions. Statistical analysis was done using one way analysis of variance, groups of 20–25 oocytes. Top inset: Western blot showing expression of all transporters in oocytes and absence of expression in water-injected oocytes. Antibody was mouse monoclonal anti-cmyc.

Article Snippet: For Western blot analysis, groups of 8 oocytes were homogenized in CHAPS-containing lysis buffer, subjected to 7.5% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), transferred to nitrocellulose membrane, incubated with mouse monoclonal anti-cmyc antibody (clone 9E10, Thermo Fisher Scientific) followed by horseradish peroxidase (HRP)-conjugated anti-mouse secondary antibody.

Techniques: Injection, Incubation, Western Blot, Expressing