Insulin is a hormone that has multiple functions including 
the stimulation of glucose transport and utilization, the 
uptake of amino acids, and the maintenance of 
differentiation. It has often been found to increase the 
growth of cultured cells, especially hybridomas. ATCC's 
Hybri-Care Medium (ATCC Cat. No. 46-X, 47-X) contains 
insulin and oxalacetic acid and will support the growth of 
80% to 90% of the hybridoma cell lines distributed.

Mechanical freezers can be used for storage if temperatures 
of -135?C or lower can be maintained. Viability will 
decline at higher temperatures. Backup freezers are needed 
in case of mechanical failure. 

Pyruvate is an intermediary organic acid metabolite in 
glycolysis and the first of the Embden Myerhoff pathway 
that can pass readily into or out of the cell. Thus, its 
addition to tissue culture medium provides both an energy 
source and a carbon skeleton for anabolic processes. Its 
addition may help in maintaining certain specialized cells, 
may help when cloning and may be necessary when the serum 
concentration is reduced in the medium (Culture of Animal 
Cells

The passage number simply refers to the number of times the 
cells in the culture have been subcultured, often without 
consideration of the inoculation densities or recoveries 
involved. The population doubling level refers to the total 
number of times the cells in the population have doubled 
since their primary isolation in vitro.

While the levels of carbon dioxide in cell culture systems 
vary from that in ambient air (about 0.03%) up to 40% in 
air, generally either no added CO2 or 5% to 10% CO2 in air 
are the most frequently used.

ATCC propagates most hybridomas as cell cultures in vitro, 
but many can be grown efficiently as ascites by inoculation 
into the peritoneal cavities of mice. Some rat-mouse 
hybridomas are more problematical as the host mouse strain 
may develop anti-rat antibodies and eventually reject the 
hybridoma inoculum. To minimize this difficulty one can use 
athymic nude mice as hosts or immunosuppress host animals 
prior to inoculation

colony forming efficiency, refers to the percentage of 
cells planted which give rise to colonies. When a cell line 
can be grown from single cells, the plating efficiency is 
determined at ATCC by inoculation of dishes or flasks with 
a suitable number of cells (100 to 1,000 cells/75 sq. cm 
flask).

use a BLASTX search against a known protein database - such 
MCBI's NR database. I would then sort them to determine how 
many were unique.

Right Angle Triangle

0.59 gms of NAOH will react with 0.2 equivalent of HCL.