3G5

Code: 0015
Cell Line: 3G5
Species: Mus musculus (B cell); Mus musculus (myeloma), mouse (B cell); mouse (myeloma)

Vulgar Name: Mouse
Morphology: Lymphoblast
Disease: Insulinoma
Growth Properties: Suspension
Derivation: Spleen cells were fused with myeloma cells.
Applications: The antibody reacts with an acetylated ganglioside common to pancreatic islet cells, neurons, adrenal medulla, thyroid follicle renal glomerulus, human T lymphocytes and a majority of cells in the pituitary. The antibody reacts with an O-acetylated disialoganglioside present on a wide variety of endocrine and neuronal tissues. The 3G5 antigen is expressed on microvascular pericyte plasma membranes, and is useful as a marker of pericytes in vitro.
Products: immunoglobulin; monoclonal antibody; against a ganglioside associated with endocrine cells (pancreatic islet, adrenal medulla, pituitary, thyroid), human T lymphocytes and neuronal cells
Biosafey: 1
Additional info: Animals were immunized with dengue virus type 2 antigens from infected mouse brains (the prototype strain New Guinea C was used). The antibody is type specific. Spleen cells were fused with P3X63Ag8 myeloma cells. Antibodies should be prepared as ascites in pristane primed BALB/c mice.
Culture Medium: Dulbecco's Modified Eagle's Medium (DMEM) modified to contain 4 mM L-glutamine, 4500 mg/L glucose, 1 mM sodium pyruvate, and 1500 mg/L sodium bicarbonate and fetal bovine serum to a final concentration of 10%.
Subculturing: Cultures can be maintained by addition of fresh medium. Alternatively, cultures can be established by centrifugation with subsequent resuspension at 1 x 10e5 viable cells/mL. Maintain cultures at a cell concentration between 1 x 10e5 and 1 x 10e6 cells/mL. NOTE: Do not allow the cell concentration to exceed 1 x 10e6 cells/mL.

Medium Renewal: Every 2 to 3 days

Culture Conditions: Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37°C
Cryopreservation: 95% FBS + 5% DMSO (Dimethyl sulfoxide)
Thawing Frozen Cells: SAFETY PRECAUTION: Is highly recommend that protective gloves and clothing always be used and a full face mask always be worn when handling frozen vials. It is important to note that some vials leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vessel exploding or blowing off its cap with dangerous force creating flying debris. 1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the Oring and cap out of the water. Thawing should be rapid (approximately 2 minutes). 2. Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions. 3. For cells that are sensitive to DMSO is recommended that the cryoprotective agent be removed immediately. Transfer the vial contents to a centrifuge tube containing 9.0 mL complete culture medium and spin at approximately 125 x g for 5 to 7 minutes. 4.Discard the supernatant and Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio). 5. Incubate the culture in a appropriate atmosphere and temperature (see "Culture Conditions" for this cell line). NOTE: It is important to avoid excessive alkalinity of the medium during recovery of the cells. It is suggested that, prior to the addition of the vial contents, the culture vessel containing the growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6).
References: Rabinowe SL, et al. Aging in man: Linear increase of a novel T cell subset defined by antiganglioside monoclonal antibody 3G5. J. Exp. Med. 165: 1436-1441, 1987. PubMed: 3494809 Powers AC, et al. Characterization of monoclonal antibody 3G5 and utilization of this antibody to immobilize pancreatic islet cell gangliosides in a solid phase radioassay. Endocrinology 114: 1338-1343, 1984. PubMed: 6368201 Nayak RC, et al. A monoclonal antibody (3G5)-defined ganglioside antigen is expressed on the cell surface of microvascular pericytes. J. Exp. Med. 167: 1003-1015, 1988. PubMed: 3351433 Bartholomeusz RK, et al. Pancreatic islet A2B5- and 3G5-reactive gangliosides are markers of differentiation in rat insulinoma cells. Endocrinology 124: 2680-2685, 1989. PubMed: 2541995
Depositors: Antonio Martins Monteiro - Banco De Células Do Rio De Janeiro
ATCC: CRL-1814