Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
This cell line is useful to study cell apoptosis, BCL-2 function and its pathway, as well as phosphorylation of BCL-2.
The BCL2 (AAA) Jurkat cell line was derived by transfecting human Jurkat T cells with the pSFFV-neo mammalian expression vector containing the human BCL2 mutated ORF insert and a neomycin-resistant gene (Parental vector: Addgene # 3349). BCL2 (AAA) stable neomycin (Neo)-resistant single-cell-originated clones were isolated and expanded by selection in medium containing 1 mg/mL G418 for 14 days
The BCL2 (AAA) Jurkat cell line was derived by transfecting human Jurkat T cells with the pSFFV-neo mammalian expression vector containing the human BCL2 mutated ORF insert and a neomycin-resistant gene (Parental vector: Addgene # 3349). The BCL2 (AAA) Jurkat cells contain the BCL2 ORF with a triple sub-mutation at Thr69, Ser70, and Ser87. These amino acids have been replaced with Alanine at their respective positions. BCL2 (AAA) stable neomycin (Neo)-resistant single-cell-originated clones were isolated and expanded by selection in medium containing 1 mg/mL G418 for 14 days. The BCL2 (AAA) Jurkat cell line overexpresses the AA/A-mutated form of Bcl-2.
Cultures can be maintained by the addition of fresh medium or replacement of medium. Alternatively, cultures can be established by centrifugation with subsequent resuspension at 1 x 105 viable cells/mL. Do not allow the cell density to exceed 3 x 106 cells/mL.
Subcultivation ratio: Maintain cultures at a cell concentration between between 1 x 105 and 1 x 106 viable cells/mL.Yamamoto K, et al. BCL-2 is Phosphorylated and Inactivated by an ASK1/Jun N-Terminal Protein Kinase Pathway normally Activated at G2/M. Mol Cell Biol. Dec;19(12):8469-78. 1999. PubMed: 10567572
Bassik MC, et al. Phosphorylation of BCL-2 regulates ER Ca2+ homeostasis and apoptosis. Embo J. March 10; 23(5): 1207–1216. 2004. PubMed: 15010700
