Quantification of telomerase activity in human liver tissues by fluorescence-based TRAP analysis

Green tea extracts attenuate doxorubicin-induced spermatogenic disorders in conjunction with higher telomerase activity in mice

“…Fluorescence-based telomeric repeat amplification protocol assay A quantitative modification of the telomeric repeat amplification protocol (TRAP) method was applied using a TRAP-eze TM telomerase detection kit (Oncor Inc.-Kyowa Co., Tokyo, Japan) according to the manufacturer's instructions [26,27]. Briefly, frozen testicular samples (20 mg for each assay) were homogenized in 100 µl of ice-cold CHAPS lysis buffer (TRAP-eze TM Telomerase Detection Kit; Oncor Inc.-Kyowa Co., Tokyo, Japan) and were incubated for 30 min on ice.…”

“…Each peak was quantified in terms of size, peak height, and peak area. Telomerase activity was quantified according to the following formula: TPG (total product generated) units/μg protein=(A×B −1 /C×D −1 ) × 100, where A is the measured total area of telomerase activity (50, 56 bp…), B is the measured area of the internal control (36 bp), C is the measured total area of telomerase activity (50, 56 bp…) in the positive control, and D is the measured area of internal control (36 bp) in the positive control [26].…”

“…Thus, we assayed telomerase activity as a functional parameter in germ cells. The detection of telomerase activity using a fluorescence-based telomeric repeat amplification protocol (F-TRAP) assay is a highly quantitative approach that has been shown to be sufficiently sensitive [26,27]. Such a system employs fluorescence-labeled primers and an automatic DNA sequencer to detect and quantify telomerase activity, which is represented by fluorocurves, with the height and area of each peak calculated automatically.…”

Regression of Gastric Mucosa-Associated Lymphoid Tissue Lymphoma with Reduced Telomerase Activity after Eradication of Helicobacter pylori.

“…Immunophenotyping (B-cell/T-cell) and immunogloblin light chain restriction were assessed using the following monoclonal antibodies; UCHL1, L26, kappa and lambda (DAKO, Carpinteria, CA, USA). Telomerase activity was measured by the TRAPmethod with modifications, using a commercially available TRAP-ezeá" Telomerase Detection Kit (ONCOR, Inc., Gaithersburg, MD, USA)as previously reported by us (27). An aliquot of a cell extract containing 1.0 [ig of protein was used for each TRAP assay.…”

“…An aliquot of a cell extract containing 1.0 [ig of protein was used for each TRAP assay. Thetelomerase activity was calculated in units of total product generated (TPG) (27,28). The results are given as the mean in TPGunits/|ig protein.…”