Assessment of the Alamar Blue assay for cellular growth and viability in vitro

Nakayama, Grace R; Caton, Maureen C.; Nova, Michael P.; Parandoosh, Zahra

doi:10.1016/s0022-1759(97)00043-4

Cited by 555 publications

(348 citation statements)

References 10 publications

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“…Viral stocks were produced in 96-well plate format and used at an arbitrary titer that was sufficient, in pilot experiments, to infect all the cells in 96-well plates containing puromycin-sensitive HeLa and 293T cells as test lines (see Methods). The renal carcinoma cells were assayed for viability 5 days after viral transduction using alamarBlue, which measures the number of metabolically active cells (26). In duplicate independent screens we observed that the raw alamarBlue values were highly reproducible (correlation coefficients Ͼ 0.90; Fig.…”

Section: -O and Rcc4 Are Clear Cell Renal Carcinoma Lines In Whichmentioning

confidence: 91%

Kinase requirements in human cells: III. Altered kinase requirements inVHL−/− cancer cells detected in a pilot synthetic lethal screen

Bommi-Reddy

Almeciga

Sawyer

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

137

117

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Clear cell renal carcinomas are the most common form of kidney cancer and frequently are linked to biallelic inactivation of the von Hippel-Lindau (VHL) tumor suppressor gene. The VHL gene product, pVHL, has multiple functions including directing the polyubiquitylation of the HIF transcription factor. We screened 100 shRNA vectors, directed against 88 kinases, for their ability to inhibit the viability of VHL؊/؊ renal carcinoma cells preferentially compared with isogenic cells in which pVHL function was restored. shRNAs for ''hits'' identified in the primary screen were interrogated in secondary screens that included shRNA titration studies. Multiple shRNAs against CDK6, MET, and MAP2K1 (also known as MEK1) preferentially inhibited the viability of 786-O and RCC4 VHL؊/؊ cells compared with their wild-type pVHL-reconstituted counterparts. The sensitivity of pVHL-proficient cells to these shRNAs was not restored upon HIF activation, suggesting that loss of an hypoxia-inducible factor (HIF)-independent pVHL function formed the basis for selectivity. A small-molecule Cdk4/6 inhibitor displayed enhanced activity against VHL؊/؊ renal carcinoma cells, suggesting that in some cases hits from shRNA screens such as described here might translate into therapeutic targets.essential kinases ͉ shRNA screens ͉ VHL ͉ kidney cancer ͉ therapeutics

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Section: -O and Rcc4 Are Clear Cell Renal Carcinoma Lines In Whichmentioning

confidence: 91%

Kinase requirements in human cells: III. Altered kinase requirements inVHL−/− cancer cells detected in a pilot synthetic lethal screen

Bommi-Reddy

Almeciga

Sawyer

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

137

117

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“…Alamar Blue dye was purchased from Biosource International Inc. (Camarillo, CA, USA) and the manufacturer's instructions were followed to complete the assay (Ahmed et al, 1994;Nakayama et al, 1997).…”

Section: Cell Proliferation (Alamarblue Assay)mentioning

confidence: 99%

Inducible re-expression of p16 in an orthotopic mouse model of pancreatic cancer inhibits lymphangiogenesis and lymphatic metastasis

et al. 2008

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Functional inactivation of the tumour suppressor protein p16 INK4a constitutes a key event in the multistep process of pancreatic ductal cell transformation. However, the significance of p16 inactivation for complex and tissue-specific aspects of pancreatic cancer progression, such as angiogenesis and metastasis, is less understood. Here, we inducibly re-expressed p16 in vivo in an orthotopic model of pancreatic cancer and examined the impact on these clinically relevant aspects of pancreatic cancer tumour biology. Consistent with previous work in subcutaneous xenograft models, we found p16 capable of reducing primary tumour growth. In addition, p16 restitution resulted in a marked reduction of tumour angiogenesis, largely accounted for by a p16-dependent inhibition of lymphangiogenesis. In excellent agreement with the antilymphangiogenic effect, re-expression of p16 almost completely prevented lymph node metastases of MiaPaca-2 pancreatic tumours. To our knowledge, this is the first report that experimentally links the tumour suppressor p16 to the process of lymphangiogenesis.

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“…The Alamar blue assay is a fluorimetric/colorimetric test based on the detection of metabolic activity [25]. It incorporates an oxidation-reduction indicator which changes from the oxidised form -non-fluorescent, blue -to the reduced state -fluorescent, red -in response to chemical reduction of growth medium, as a result of cell growth.…”

Section: Toxicity and Cell Growth Inhibition Evaluationmentioning

confidence: 99%

Inclusion of molybdenocene dichloride (Cp2MoCl2) in 2-hydroxypropyl- and trimethyl-β-cyclodextrin: Structural and biological properties

Braga

Marques

Sousa

et al. 2005

Journal of Organometallic Chemistry

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Organometallic-cyclodextrin inclusion compounds were obtained by the treatment of molybdenocene dichloride (Cp 2 MoCl 2 ) with the modified cyclodextrins (CDs) heptakis-2,3,6-tri-O-methyl-b-CD (TRIMEB) and 2-hydroxypropyl-b-CD (HPbCD) in aqueous solution. The products were isolated by liophilisation and characterised in the solid-state by powder XRD, thermogravimetric analysis, Raman and FTIR spectroscopy, and 13 C CP MAS NMR spectroscopy. The results are consistent with inclusion of Cp 2 MoCl 2 , rather than hydrolysis products such as [Cp 2 Mo(H 2 O)X] + (X = Cl, OH) or [Cp 2 Mo(H 2 O) 2 ] 2+ . The pure non-included metallocene Cp 2 MoCl 2 and its inclusion compounds with unmodified b-CD, TRIMEB and HPbCD were screened for their potential antiproliferative and cytotoxic activity, in both human cancer and healthy cell lines. Inclusion in CD was found to enhance the cytotoxic effect of Cp 2 MoCl 2 , with the TRIMEB adduct displaying the highest anti-tumour activity, along with the lowest toxicity towards non-neoplastic cells.

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Assessment of the Alamar Blue assay for cellular growth and viability in vitro

Cited by 555 publications

References 10 publications

Kinase requirements in human cells: III. Altered kinase requirements inVHL−/− cancer cells detected in a pilot synthetic lethal screen

Kinase requirements in human cells: III. Altered kinase requirements inVHL−/− cancer cells detected in a pilot synthetic lethal screen

Inducible re-expression of p16 in an orthotopic mouse model of pancreatic cancer inhibits lymphangiogenesis and lymphatic metastasis

Inclusion of molybdenocene dichloride (Cp2MoCl2) in 2-hydroxypropyl- and trimethyl-β-cyclodextrin: Structural and biological properties

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