Cell Culture Reagents, Cell Culture Reagents
Trypan Blue, 0.4% Solution In PBS
Trypan Blue is a blue acid dye with a strong affinity for cellulose containing substrates such as cotton; less affinity for proteinaceous materials. Trypan blue solution may be used in trypan blue based cytotoxitiy and proliferation assays. Trypan Blue is used as a vital dye which is especially important because it is taken up by the reticuloendothelial system. Clark describes assays for the study of teratogenic action of trypan blue on embryonic tissues using Davis and Sauter's fluorescence method and for the staining of collagen, including very fine fibrils, muscle and cornified epithelium using the Van Gieson method. Trypan blue is also recommended for use in dye exclusion procedures for viable cell counting. Non-viable cells will up-take trypan blue at a faster rate than viable cells.
Endothelial Cell Growth Supplement
From Bovine Hypothalamus
Lyophilized from sodium chloride buffer containing streptomycin sulfate
Activity: promotes the growth of Fetal Bovine Heart Endothelial Cells (FBHEC) in 10% newborn calf serum when reconstituted at a concentration of 50-150 μg/ml serum-free medium.
It contains several growth factors (e.g. FGF, ECGF-α) and supports the growth of vascular endothelial cells from human and other mammalian species. Also, it may be used for the cloning and proliferation of hybridomas and human keratinocytes.
Each lot is sterile filtered and tested for bacteria, fungi, and mycoplasma contamination. Some cells may require supplementation with 1-100 μg/ml heparin.
Epidermal Growth Factor 95%
Epidermal growth factor or EGF is a growth factor isolated from mouse submaxillary glands that stimulates cell growth, proliferation, and differentiation by binding to its receptor EGFR. EGF is mitogenic for a variety of epidermal and epithelial cells, including fibroblasts, glial cells, mammary epithelial cells, vascular and corneal endothelial cells, bovine granulosa, rabbit chondrocytes, HeLa and SV40-3T3 cells.
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.
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.
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).
The antibiotics penicillin and streptomycin are used to prevent bacterial contamination of cell cultures due to their effective combined action against gram-positive and gram-negative bacteria. This solution contains 10,000 units/mL of penicillin and 10,000 µg/mL of streptomycin.
Amphotericin-B (250 µg/mL) 20 mL
Amphotericin B is a polyene antifungal antibiotic from Streptomyces sp. It has a high affinity for sterols, primarily ergosterols, of fungal and bacterial cell membranes. It is an antibiotic solution effective against yeast and other fungi. Amphotericin B may be toxic to some insect cell types. Amphotericin B, is an antimycotic.