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-100°C or colder, preferably in the vapor phase of a liquid nitrogen freezer
Quantity limit per order for this item is 1. This item can be ordered twice a year. Orders over this limit will be sent to NIAID for approval before shipment.
ARP-12197 is an Epstein–Barr virus-transformed Macaca mulatta B cell that produces the IgG1λ monoclonal antibody 4B2. It has not been fused with any myeloma cell line.
4B2 binds the highly immunogenic region at the beginning of the cytoplasmic tail of Simian immunodeficiency virus (SIVmac) gp41 (also known as the Kennedy Epitope). It was isolated from a SIVmac239-infected monkey.
Antibody secretion is continuous but is optimal about two weeks after transferring to serum-free media or when the medium begins to turn yellow. The expected antibody yield is > 1 mg per liter of serum-free medium.
The monoclonal antibody binds the original peptide used for screening (CFSSPPSYFQQTHIQQDPALPTREGKER) but not two shorter versions (QQTHIQQDPALPTRE and IQQDPALPTREGKER). The minimum positive peptide tested is: CFSSPPSYFQQTHIQQDPALPTREGKER. The epitope overlaps with monoclonal antibody 4B2 (ARP-12224, produced by ARP-12196). The antibody does not bind by ELISA to recombinant protein SIVmac239 gp120 or gp140 (both lack the domain responsible for binding) or to any other of the peptides overlapping the whole SIV envelope glycoprotein (Env). There is no detectable neutralization for the tested viruses.
Growth characteristics: After thawing the vial and washing the freezing media away, it is advised to seed the cells in 10 mL of media in a T75 in the vertical position. Add fresh media when the original media yellows and expand to the desired amount. Split the culture no higher than 1:4. Cells grow mostly in suspension, singly and in small clumps, with some weakly adhering to the flask, and appear brilliant and round with a few small membrane protrusions under magnification. Propagation Medium: RPMI 1640 medium supplemented with 15% fetal bovine serum, L-glutamine, non-essential amino acids, sodium pyruvate, HEPES and Primocin™ (InvivoGen).
ARP-12197 is negative for Mycoplasma, bacteria and fungi. Each vial of ARP-12197 contains approximately 6 x 106 cells in fetal bovine serum containing 10% dimethyl sulfoxide (DMSO). Please refer to the appropriate data sheet for lot-specific information.
Postler, T. S., et al. “Evidence Against Extracellular Exposure of a Highly Immunogenic Region in the C-Terminal Domain of the Simian Immunodeficiency Virus gp41 Transmembrane Protein.” J. Virol. 86 (2012): 1145-1157. PMID: 22072749.
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