aCella - Tox - an assay designed to measure cell cytoxicity

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A bioluminescence, non-radioactive cytotoxicity assay designed to quantitatively measure the release of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from mammalian cell lines or bacterial cells.

5000 Ramos cells/well were incubated with serially diluted Rituxan antibody for 15 minutes prior to the addition of purified NK cells stimulated overnight with IL-2. The ADCC reaction was further incubated for 2 hours at the specified E:T ratios. % Cytotoxicity was measured using the aCellaTOX assay.
5000 Ramos cells/well were incubated with serially diluted Rituxan antibody for 15 minutes prior to the addition of purified NK cells stimulated overnight with IL-2 . The ADCC reaction was further incubated for 3.5 hours at the specified E:T ratios. % Cytotoxicity was measured using the aCellaTOX assay. 3.5hour timepoint has been optimized with the 10:1 E:T ratio.
The graph shows an antibody titration of Rituximab from0.000977 to 1ug/ml incubated with 5% baby rabbit complement and 5000 Daudi target cells for 30 minutes at 37oC
5000 Daudi cells/well were incubated with serially diluted Rituxan antibody for 15 minutes prior to the addition of purified NK cells stimulated overnight with IL-2. The ADCC reaction was further incubated for 2 hours at the specified E:T ratios. % ADCC was measured using the aCella-TOX assay.Log (EC50) value for E:T 25:1 was -2.344 and for E:T 10:1 was -2.213
A Direct comparison of 51Cr and aCella-TOX was carried out with the same donors with Daudi Cells. Log (EC50) value for aCella-TOX was -2.23, and that for 51Cr was -2.087 for Donor A.
A Direct comparison of 51Cr and aCella-TOX was carried out with the same donors with Daudi Cells. Log(EC50) value for aCella-TOX was -2.612, and that for51Cr was -2.77 for Donor B.
5000 Ramos cells/well were incubated with serially diluted Rituxan antibody for 15 minutes prior to the addition of purified NK cells stimulated overnight with IL-2. The ADCC reaction was further incubated for 2 hours at the specified E:T ratios. % Cytotoxicity was measured using the aCellaTOX assay.
5000 Ramos cells/well were incubated with serially diluted Rituxan antibody for 15 minutes prior to the addition of purified NK cells stimulated overnight with IL-2. The ADCC reaction was further incubated for 2 hours at the specified E:T ratios. % Cytotoxicity was measured using the aCellaTOX assay.
5000 Ramos cells/well were incubated with serially diluted Rituxan antibody for 15 minutes prior to the addition of purified NK cells stimulated overnight with IL-2 . The ADCC reaction was further incubated for 3.5 hours at the specified E:T ratios. % Cytotoxicity was measured using the aCellaTOX assay. 3.5hour timepoint has been optimized with the 10:1 E:T ratio.
The graph shows an antibody titration of Rituximab from0.000977 to 1ug/ml incubated with 5% baby rabbit complement and 5000 Daudi target cells for 30 minutes at 37oC
5000 Daudi cells/well were incubated with serially diluted Rituxan antibody for 15 minutes prior to the addition of purified NK cells stimulated overnight with IL-2. The ADCC reaction was further incubated for 2 hours at the specified E:T ratios. % ADCC was measured using the aCella-TOX assay.Log (EC50) value for E:T 25:1 was -2.344 and for E:T 10:1 was -2.213
A Direct comparison of 51Cr and aCella-TOX was carried out with the same donors with Daudi Cells. Log (EC50) value for aCella-TOX was -2.23, and that for 51Cr was -2.087 for Donor A.
A Direct comparison of 51Cr and aCella-TOX was carried out with the same donors with Daudi Cells. Log(EC50) value for aCella-TOX was -2.612, and that for51Cr was -2.77 for Donor B.

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Data Sheet
Protocol
Safety Data Sheet

SKU: CLATOX100-4

Size: 1000 Tests
Price:
$1,703.00
GAPDH is an important enzyme in the glycolysis and gluconeogenesis pathways. This homotetrameric enzyme catalyzes the oxidative phosphorylation of D-glyceraldehyde-3-phosphate to 1,3-diphosphoglycerate in the presence of cofactor and inorganic phosphate. In the aCella-TOX reaction scheme the release of GAPDH is coupled to the activity of the enzyme 3-Phosphoglyceric Phosphokinase (PGK) to produce ATP. ATP is detected via the luciferase, luciferin Bioluminescence methodology. Further, aCella-TOX is a homogeneous cytotoxicity assay; alternatively in dual mode, aCella-TOX can measure cytotoxicity and cell viability in the same plate. Culture supernatants can also be removed from the original plate and assayed in a different plate, allowing kinetics runs to be set up. The assay is non-destructive, allowing the monitoring of additional parameters such as gene expression.
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CLATOX100-3P: 500 Tests (with 5 Lumi Plates + 5 TC Plates)
  • 1. 4X Enzyme Assay Reagent Part# 6001 x 1 (Storage: -20°C)
  • 2. Enzyme Assay Diluent Part# 3008 x 1 (Storage: 2-8°C)
  • 3. 50X Detection Reagent Part# 6002 x 1 (Storage: -20°C)
  • 4. 5.5X Detection Assay Diluent Part# 3009 x 1 (Storage: -20°C)
  • 5. Glyeraldehyde 3-Phosphate-G3P Part# 6003 x 1 (Storage: -20°C)
  • 6. Lysis Buffer Part# 3035 x 1 (Storage: 2-8°C)
  • 7. 5 Luminescence Plates + 5 Tissue Culture Plates (Catalog# CLATOX100-3P)

    • CLATOX100-3: 500 Tests
  • 1. 4X Enzyme Assay Reagent Part# 6001 x 1 (Storage: -20°C)
  • 2. Enzyme Assay Diluent Part# 3008 x 1 (Storage: 2-8°C)
  • 3. 50X Detection Reagent Part# 6002 x 1 (Storage: -20°C)
  • 4. 5.5X Detection Assay Diluent Part# 3009 x 1 (Storage: -20°C)
  • 5. Glyeraldehyde 3-Phosphate-G3P Part# 6003 x 1 (Storage: -20°C)
  • 6. Lysis Buffer Part# 3035 x 1 (Storage: 2-8°C)

    • CLATOX100-4: 1000 Tests
  • 1. 4X Enzyme Assay Reagent Part# 6001 x 2 (Storage: -20°C)
  • 2. Enzyme Assay Diluent Part# 3008 x 2 (Storage: 2-8°C)
  • 3. 50X Detection Reagent Part# 6002 x 2 (Storage: -20°C)
  • 4. 5.5X Detection Assay Diluent Part# 3009 x 2 (Storage: -20°C)
  • 5. Glyeraldehyde 3-Phosphate-G3P Part# 6003 x 2 (Storage: -20°C)
  • 6. Lysis Buffer Part# 3035 x 2 (Storage: 2-8°C)

    • CLATOX100-3L: 500 Tests (with 5 Lumi Plates)
  • 1. 4X Enzyme Assay Reagent Part# 6001 x 1 (Storage: -20°C)
  • 2. Enzyme Assay Diluent Part# 3008 x 1 (Storage: 2-8°C)
  • 3. 50X Detection Reagent Part# 6002 x 1 (Storage: -20°C)
  • 4. 5.5X Detection Assay Diluent Part# 3009 x 1 (Storage: -20°C)
  • 5. Glyeraldehyde 3-Phosphate-G3P Part# 6003 x 1 (Storage: -20°C)
  • 6. Lysis Buffer Part# 3035 x 1 (Storage: 2-8°C)
  • 7. 5 Luminescence Plates (Catalog# CLATOX100-3L)

    • Product Specific References

    PMID Publication
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    30867324Sano, R., et al. 2019. An antibody-drug conjugate directed to the ALK receptor demonstrates efficacy in preclinical models of neuroblastoma. Science Translational Medicine, eaau9732.
    31070879Riss, T., et al. 2019. Cytotoxicity assays: in vitro methods to measure dead cells. Assay Guidance Manual [Internet]. Bethesda (MD): Eli Lilly & Company and the National Center for Advancing Translational Sciences, .
    25391374Sanabria-Figueroa, E., et al. 2015. Insulin-like growth factor-1 receptor signaling increases the invasive potential of human epidermal growth factor receptor 2-overexpressing breast cancer cells via Src-focal adhesion kinase and forkhead box protein M1. Molecular Pharmacology , 150-61.
    25915748Simhadri, V.R., et al. 2015. A Human Anti-M2 Antibody Mediates Antibody-Dependent Cell-Mediated Cytotoxicity (ADCC) and Cytokine Secretion by Resting and Cytokine-Preactivated Natural Killer (NK) Cells. PLoS One, e0124677.
    30804020Bekaii-Saab, T.., et al. 2019. Phase I Immunotherapy Trial with Two Chimeric HER-2 B-Cell Peptide Vaccines Emulsified in Montanide ISA 720VG and Nor-MDP Adjuvant in Patients with Advanced Solid Tumors. Clinical Trial Clin Cancer Res, 3495-3507.
    25932028Tsuji, W., et al. 2015. Effects of immunosuppressive drugs on viability and susceptibility of adipose-and bone marrow-derived mesenchymal stem cells. Frontiers in Immunology, 131.
    26431358Fisher, T.L., et al. 2016. Generation and preclinical characterization of an antibody specific for SEMA4D. Mabs, 150-62.
    26390038Liu, J., et al. 2015. Pre-clinical development of a humanized anti-CD47 antibody with anti-cancer therapeutic potential. PLoS One, e0137345.
    34395257Lewis, C.S., et al. 2021. A First-In-Class, Humanized Antibody Targeting Alternatively Spliced Tissue Factor: Preclinical Evaluation in an Orthotopic Model of Pancreatic Ductal Adenocarcinoma. Frontiers in Oncology, 691685.
    36211807Guo, L., et al. 2022. A newly discovered PD-L1 B-cell epitope peptide vaccine (PDL1-Vaxx) exhibits potent immune responses and effective anti-tumor immunity in multiple syngeneic mice models and (synergizes) in combination with a dual HER-2 B-cell vaccine (B-Vaxx). Oncoimmunology, 2127691.
    25542973Zhukovskaya, N.L., et al. 2015. The processivity factor complex of feline herpes virus-1 is a new drug target. Antiviral Research, 17-20.
    32307580Zecca, A., et al. 2020. Energy metabolism and cell motility defect in NK-cells from patients with hepatocellular carcinoma. Cancer Immunology, Immunotherapy, 1589-1603.
    34895654Láng, J.A., et al. 2020. In vitro functional characterization of biosimilar therapeutic antibodies. Drug Discovery Today: Technologies, 41-50.
    28970093Hoffman, F., et al. 2018. A mathematical model of antibody-dependent cellular cytotoxicity (ADCC). Journal of Theoretical Biology, 39-50.
    34931298Dash, R., et al. 2022. Assessment of Functional Characterization and Comparability of Biotherapeutics: a Review. AAPS Journal, 15.

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