alamarBlue® references


alamarBlue® - Cell Proliferation and Viability Reagent

alamarBlue - Cell Proliferation and Viability Reagent

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Listed here are alamarBlue articles and educational resource for anyone interested in learning more about the detailed methods, uses and advantages of alamarBlue.

Comparisons with alternative cell proliferation assays such as MTT, XTT and [3H]thymidine incorporation assays have been highlighted.

Articles have also been grouped by methods to make it easier to see how alamarBlue is used in a variety of cell culture applications. 

Comparison of alamarBlue to MTT assays

It has been shown by Hamid et al. in 2004, that alamarBlue is more sensitive than 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) for most of the drug compounds they tested in HepG2 cell viability assays.

  • Hamid, R. et al. (2004). Comparison of alamar blue and MTT assays for high through-put screening.
    Toxicol In Vitro. 18(5):703-10
  • Alley, M.C. et al. (1988). Feasibility of Drug Screening with Panels of Human Tumor Cell Lines Using a Microculture Tetrazolium Assay.
    Cancer Res. 48: 589-601 

Comparison of alamarBlue to XTT assays

  • Pagé et al. in 1993 describe how alamarBlue showed comparable results to the XTT formazan assay, that alamarBlue was more economical and faster at producing results. It was also highlighted that MTT and XTT are low in sensitivity, expensive and harmful reagents compared to alamarBlue.
  • Pagé, B. et al. (1993). A new Fluorometric Assay for Cytotoxicity Measurements In Vitro.
    Int. J. Oncology 3: 473-476

Comparison of alamarBlue to [3H]thymidine incorporation assay

A study on the use of alamarBlue with peripheral blood mononuclear cells (PBMC) by De Fries et al. 1995 showed that the alamarBlue assay reliably detects human PBMC, and the results were highly reproducible. Ahmed et al. in 1994 describe the use of alamarBlue to monitor and determine the proliferation of murine lymphocytes, lymphoid tumor and hybridoma cells. Cell proliferation can be determined by color change using an ELISA plate reader after alamarBlue is added during the initial phase of cell culture.  It was determined that alamarBlue gave comparable results to the [3H]thymidine incorporation assay. 

The summary of alamarBlue advantages over the [3H]thymidine incorporation assay include:

  • Non-radioactive
  • Simplicity
  • Less costly
  • Non-labor intensive
  • rapidity of assessment of proliferation of large number of samples
  • non-toxicity
  • usefulness in determining the kinetics of cell growth of hybridomas
  • non-interference of secretion of antibodies by a hybridoma cell line
  • Ahmed, S.A. et al. (1994). A new Rapid and Simple Non-Radioactive assay to Monitor and Determine the proliferation of Lymphocytes: An Alternative to H3-thymidine incorporation assay.
    J. Immunol. Methods.170:211-24
  • De Fries, R. et al. (1995). Quantification of Mitogen Induced Human Lymphocyte Proliferation: Comparison of alamarBlueTM to 3H-Thymidine Incorporation Assay.
    J. Clin. Lab. Anal. 9: 89-95 

Cell proliferation and viability assay articles

  • Breinholt, V. et al. (1998). Detection of Weak Estrogenic Flavonoids Using a Recombinant Yeast Strain and a Modified MCF7 Cell Proliferation Assay.
    Chem. Res. Toxicol. 11: 622-629
  • Foresti, R. et al. (2005). Differential Activation of Heme Oxygenase-1 by Chalcones and Rosolic Acid in Endothelial Cells.
    J. Pharmacol. Exp. Ther. 312: 686-693
  • Choi, J. et al. (2005). CD137 Induces Adhesion and Cytokine Production in Human Monocytic THP-1 cells.
    Exp.Mol. Med. 37: 78-85
  • Barbero, A. et al. (2005). Experimental and Mathematical Study of the Influence of Growth Factors on the Growth Kinetics of Adult Human Articular Chondrocytes.
    J. Cell. Physiol. 204: 830-838
  • Adikari, S.B. et al. (2004). Interferon-modified Dendritic Cells Suppress B Cell Function and Ameliorate the Development of Experimental Autoimmune Myasthenia Gravis.
    Clin. Exp. Immunol. 138: 230-236
  • Giordano, C. et al. (2004). Titanium for Osteointegration: Comparison Between a Novel Biomimetic Treatment and Commercially Exploited Surfaces.
    J. Appl. Biomat. Biomech. 2: 35-44
  • Borg, S.A. et al. (2003). Expression of Interleukin-6 and its Effects on Growth of HP75 Human Pituitary Tumor Cells.
    J. Clin. Endocrinol. Metab. 88: 4938-4944
  • Dawson, C.W. et al. (2003). Epstein-Barr Virus Latent Membrane Protein 1 (LMP1) Activates the Phosphatidylinositol 3-kinase/ Akt Pathway to Promote Cell Survival and Induce Actin Filament Remodelling.
    J. Biol. Chem. 278: 3694-3704
  • Mohan, J. et al. (2005). Neuroinvasion by Scrapie following Inoculation via the Skin Is Independent of Migratory Langerhans Cells.
    J. Virol. 79: 1888-1897
  • O'Brien et al. (2000). Investigation of the Alamar Blue (resazurin) fluorescent dye for the assessment of mammalian cell cytotoxicity.
    European J. of Biochem. 267:5421-5426
  • Brieger, A. et al. (2002). Transient Mismatch Repair Gene Transfection For Functional Analysis of Genetic hMLH1 and hMSH2 Variants.
    Gut 51: 677-684
  • Scapagnini, G. et al. (2002). Caffeic Acid Phenethyl Ester and Curcumin: A Novel Class of Heme Oxygenase-1 Inducers.
    Mol. Pharmacol. 3: 554-561
  • Guo, Y. et al. (2002). An Antiangiogenic Urokinase-derived Peptide Combined with Tamoxifen Decreases Tumor Growth and Metastasis in a Syngeneic Model of Breast Cancer.
    Cancer Res. 62: 4678-4684
  • Tiwari, A. et al. (2002). Development of a Hybrid Cardiovascular Graft Using a Tissue Engineering Approach.
    FASEC J. 16: 791-796
  • Sakurai, T. et al. (2001). Modulation of Cell Adhesion and Viability of Cultured Murine Bone Marrow Cells by Arsenobetaine, a Major Organic Arsenic Compound in Marine Animals.
    Br. J. Pharmacol. 132: 143-150
  • Karsdal, M.A. et al. (2003). Transforming Growth Factor-beta Controls Human Osteoclastogenesis Through the p38 MAPK and Regulation of RANK Expression.
    J. Biol. Chem. 278: 44975-44987
  • Qureshi, M. et al. (2001). Neonatal Cells in an Adult Lung Environment are Competent to Resolve Pneumocystis carinii Pneumonia.
    J. Immunol. 166: 5704-5711
  • Yanagida, M. et al. (1995). Effects of T-helper 2-type Cytokines, Interleukin-3 (IL-3), IL-4, IL-5 and IL-6 on the Survival of Cultured Human Mast Cells.
    Blood. 86: 3705-3714
  • Yang, S. Y. et al. (2008). Inducing Apoptosis of Human Colon Cancer Cells by an IGF-I D Domain Analogue Peptide
    Mol. Cancer 7:17

Cell metabolism studies  

  • Naughton, P. et al. (2002). Induction of Heme Oxygenase 1 by Nitrosative Stress. A Role for Nitroxyl Anion.
    J. Biol. Chem. 277: 40666-40674
  • Hattori, Y. et al. (2002). Vascular Smooth Muscle Cell Activation by Glycated Albumin (Amadori Adducts).
    Hypertension. 39: 22-28
  • Park, Y.H. et al. (2000). Phenotypic and Functional Analysis of Bovine Gamma Delta Lymphocytes.
    J. Vet. Sci. 1: 39-48

Virus, drug susceptibity and toxicity studies

  • Ariyanayagam, M.R. et al. (2005). Phenotypic Analysis of Trypanothione Synthetase Knockdown in the African Trypanosome.
    Biochem J. 391: 425-32
  • Habtemariam, S. et al. (2003). In Vitro Antileishmanial Effects of Antibacterial Diterpenes from Two Ethiopian Premma Species: P. schimperi and P. oligotricha.
    BMC Pharmacology 3: 6-11
  • Casarosa, P. et al. (2003). Identification of the First Nonpeptidergic Inverse Agonist for a Constitutively Active Viral-encoded G Protein-coupled Receptor.
    J. Biol. Chem. 278: 5172 – 5178
  • Nestler, U. et al. (2002). The tissue level of dexamethasone in human brain tumors is about 1000 times lower than the cytotoxic concentration in cell culture.
    Neurol. Res. 24: 479-482
  • McCormick, A.L. et al. (2001). Immunization with an Interferon-g–gp120 Fusion Protein Induces Enhanced Immune Responses to Human Immunodeficiency Virus gp120.
    J. Infect. Dis. 184: 1423–1430
  • Latham, J.P.F. et al. (2000). Prostate-specific Antigen Promoter/Enhancer Driven Gene Therapy for Prostate Cancer: Construction and Testing a Tissue-specific Adenovirus Vector.
    Cancer Res. 60: 334-341
  • Simms, J.R. et al. (2000). Use of Herpes Simplex Virus (HSV) Type 1 ISCOMS 703 Vaccine for Prophylactic and Therapeutic Treatment of Primary and Recurrent HSV-2 Infection in Guinea Pigs.
    J. Infect. Dis. 181: 1240-1248

Microbial studies

  • Ferro, V.A. et al. (2003). In Vitro Susceptibilities of Shigella flexneri and Streptococcus pyogenes to Inner Gel of Aloe barbadenis Miller.
    Antimicrob. Agents Chemother. 47: 1137-1139
  • Pettit, R.K. et al. (2005). Microplate alarm blue assay for Staphylococcus epidermidis biofilm susceptibility testing.
    Antimicrob. Agents Chemother. 49(7): 2612 – 2617

Helpful method articles

  • Fields, R.D. and Lancaster, M.V. (1993). Dual Attribute Continuous Monitoring of Cell Proliferation/Cytotoxicity.
    Am. Biotechnol. Lab. 11: 48-50 
  • Geier, S. Ph. D (1994). Personal Communication: Analysis of alamarBlueTM Overlap: Contribution of Oxidised (ABO/OD600nm) to Reduced (ABR/OD570) OD
  • William, H.H. et al. (1965). Ultraviolet and Visible Absorption Methods, p. 94-95, in: instrumental Methods of Analysis, D. Van Nostrand Co. Inc., Princeton, N.J
  • Goegan, P. et al. (1995). Effects of Serum Protein and Colloid on the alamarBlueTM Assay in Cell Cultures.
    Toxic InVitro. 9: 257-266

Patent articles

  • Lancaster, M.V. and Fields, R.D. (1996). Antibiotic and Cytotoxic Drug Susceptibility Assays using Resazurin and Poising Agents.
    U.S. Patent No. 5,501,959