Cell Culture Reagents
Hemoglobin is the major component of erythrocytes, and is responsible for the red color. Its normal concentration in erythrocytes is approximately 34%. It is the most important respiratory protein of vertebrates by virtue of its ability to transport oxygen to tissues and carries carbon dioxide away from the tissues. Anomalous globins in which various amino acids have been substitued with others, or in which certain amino acids are missing entirely from the normal sequence, comprise 153 abnormal hemoglobin configurations. Sickle cell anemia is one such configuration. In sickle cell hemoglobin, a valine residue has replaced the glutamic acid residue at position 6 in the b-chain of normal hemoglobin; the a-chain is normal.
1M HEPES Buffer
HEPES (N-2-hydroxyethylpiperazine-N-2-ethane sulfonic acid) is a general purpose zwitterionic organic chemical buffering agent which does not bind magnesium, calcium, manganese(II) or copper (II) ions. Buffer strength for cell culture applications is usually in the range of 10 to 25 mM. Studies have indicated that 20 mM HEPES is the most satisfactory concentration of the buffer when both Hanks' and Earle's solutions are used.
Hybridoma Cell Growth Supplement
From Cultured Human Endothelial Cells
HCGS will significantly improve the hybridoma cell cloning efficiency while eliminating feeder cell need.
HCGS may be used in dilutions of 1:5 to 1:10 (v/v). We recommend HCGS be thawed and sterile filtered (0.22 or 0.45 membrane) immediately prior to use.
Interleukin-2, Human, Natural
Human Interleukin-2 (IL-2), or Human T-cell growth factor is a glycoprotein which promotes in-vitro proliferation and long-term growth of T-cell lymphokines. Additionaly, IL-2 stimulates production of cytotoxic lymphocytes, natural killer cells, and lymphokine-activated killer cells.
200 mM L-Glutamine (Frozen)
L-glutamine is an essential amino acid that is a crucial component of culture media that serves as a major energy source for cells in culture. L-glutamine is very stable as a dry powder and as a frozen solution. In liquid media or stock solutions, however, L-glutamine degrades relatively rapidly. Optimal cell performance usually requires supplementation of the media with L-glutamine prior to use.
Phenol Red, 0.5% Solution
Phenol red is an acid-base indicator. It is made by condensing two moles of phenol with one mole of o-sulfobenzoic acid anhydride. Phenol red in tissue culture media can act as a weak estrogen, especially with human breast cancer cells. Phenol red can be used to measure hydrogen peroxide in cultured macrophages in multiwell plates.
Endothelial Cell Growth Supplement
From Bovine Brain
Lyophilized powder with streptomycin sulfate.
ECGS is a broad culture medium growth supplement containing several growth factors which supports the growth of mammalian vascular endothelial cells and can be used in replacement of cell feeder layers in support of hybridoma clonal growth. It is isolated from bovine brain by a modification of the method of Maciag.
Ref.: Maciag, et al., Proc. Nat'l. Acad. Sci. USA, 76: 5674-5678 (1979).
Donor Horse Serum
This serum is processed from whole blood collected from a controlled herd of young horses. Each animal is routinely screened for Equine Infectious Anemia (EIA). Donor Horse Serum is suitable for use in diagnostic assays, as a supplement in mycoplasma growth and assay media and for culturing hematopoietic stem cells and neuronal cells.