NALM6 leukemic cells were acquired from the American Type Culture Collection (ATCC), routinely authenticated by the University of Arizona Genetics Core, and confirmed negative for Mycoplasma. NALM6 cells were transduced with a lentiviral vector encoding click beetle green (CBG) luciferase and GFP under and elongation factor 1 alpha (EF1α) promoter, separated by a P2A bicistronic linker. A fluorescence-activated cell sorter (FACS) was then used to collect ~100% GFP-positive cells, which were then expanded and cryopreserved for future use. CD19 and GFP positivity was routinely checked prior to use for in vitro and in vivo experiments by flow cytometry. NALM6 cells were maintained in culture with R10 medium: RPMI 1640 (Gibco) supplemented with 10% heat-inactivated fetal bovine serum (FBS; Seradigm), 2% 1M Hepes buffer solution (Gibco), 1% 100× GlutaMAX (Gibco), and 1% penicillin (10,000 U/mL) + streptomycin (10,000 µg/mL).
1m hepes buffer solution
Manufactured by Thermo Fisher Scientific
1M Hepes buffer solution is a commonly used buffer solution for maintaining a stable pH environment in various laboratory applications. It is an aqueous solution containing the Hepes (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) chemical compound at a concentration of 1 molar. The primary function of this buffer solution is to help maintain a consistent pH level in biological and biochemical experiments, cell culture systems, and other relevant laboratory procedures.
Automatically generated - may contain errors
2 protocols using 1m hepes buffer solution
1
Tracking NALM6 Leukemic Cells
2
Brain and Bladder Tissue Sampling
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Dissection of the brain was performed in a biosafety cabinet. Two pieces of brain tissue (approximately 5 mm  5 mm  3 mm) were taken from both the right and the left cerebral hemispheres using sterile instruments. The tissue sample from the right (irradiated) hemisphere was taken from the centre of the irradiation array and the sample from the left (unirradiated) hemisphere was taken from the corresponding contralateral location. Samples were placed in a 5 ml sterile tube containing 1 mL of Roswell Park Memorial Institute (RPMI 1640, Gibco) growth medium, supplemented with 10% FBS, 5 ml of Penicillin-Streptomycin (Gibco), 5 ml of L- glutamine (Gibco), 0.5 mg/ml of Hydrocortisone (SigmaeAldrich), and 12.5 ml of 1 M HEPES buffer solution (Gibco). Samples were immediately transported on ice to the tissue culture laboratory to be prepared for explant culture. The remaining brain tissue was snap-frozen in liquid nitrogen and stored at À80 C for proteomic studies. The entire extracted urinary bladder was also placed in a sterile 5 ml tube containing 1 ml of complete growth medium and used to set up tissue explant cultures.
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