GenomONE-CF FZ SeV-E (HVJ-E) 1 vial Cell Fusion Reagents
GenomONE™-CF EX is a cell fusion kit composed of HVJ Envelope (HVJ-E) and special buffers.
It can be used with both adhering cells and floating cells. Fusion of cells of the same or different types is possible with this kit in only 30 minutes
Feature and Advantages
Cmparison with PEG method in hybridoma preparation
hybridoma
growth supplementhybridoma
positive rateantibody-production
positive rateGenomONE - CF EX - 38/96 (40%) 9/96 (9%) + 96/96 (100%) 96/96 (100%) PEG1500 - 3/96 (3%) 1/96 (1%) + 36/96 (38%) 9/96 (9%) GenomONE - CF EX produces more antibody secreting hybridomas than PEG. (following cell fusion, cells were grown in media containing hybridoma growth supplement as indicated).
「Normal BALB/c mouse splenocytes (1x10ˆ8 cells) not sensitized with antigen were fused to X63-Ag8.653 myeloma cells (1x10ˆ7 cells) using GenomONE(tm)-CF EX or PEG1500. The fused cells obtained with each agent were inoculated onto five 96-well plates (Day 0). Beginning the following day, half of the culture medium (10%FBS/RPMI1640) was replaced with HAT medium at five points of time (Days 1, 2, 3, 5, and 8), and the growth of colonies in each well was assessed on Days 10 -11 to determine the hybridoma-positive rate
(an indicator of efficiency of fusion). On Day 12, mouse antibody level (IgG + IgA + IgM) in the supernatant was measured by ELISA, to calculate the antibody production-positive rate. The effect of adding a commercially available hybridoma supplement to the medium after fusion was also assessed (supplement was also added to the HAT medium).