alamarBlue® references


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

alamarBlue® is manufactured for Bio-Rad by Trek Diagnostic System. U.S. patent 5,501,959.