CriticalAppraisaloftheMTTAssayinthePresenceofRottlerin2

Materials and methodsMaterials All chemicals and materials for cell culture (unless otherwise indicated) were obtained from Sigma (Milan, Italy). Lactate dehydrogenase (LDH) assay kit was purchased from Sclavo Diagnostics (Siena, Italy). Rottlerin was dissolved in dimethyl sulfoxide (DMSO). Carbonylcyanide-4-(trifluoromethoxy)-phenylhydrazone (FCCP) was dissolved in 95% ethanol.Cells and culture condition......阅读全文

Critical-Appraisal-of-the-MTT-Assay-in-the-Presence-of-Rottlerin-2

Materials and methodsMaterials   All chemicals and materials for cell culture (unless otherwise indicated) were obtained from Sigma (Milan, Italy). La

Critical-Appraisal-of-the-MTT-Assay-in-the-Presence-of-Rottlerin-1

Rottlerin is a natural product isolated from Mallotus philippinensis. This polyphenolic compound, originally described as a selective inhibitor of PKC

Critical-Appraisal-of-the-MTT-Assay-in-the-Presence-of-Rottlerin-3

LDH assay   LDH assay was performed in culture medium of untreated confluent cells by using a commercial kit (Sclavo Diagnostics, Siena) based on the

Critical-Appraisal-of-the-MTT-Assay-in-the-Presence-of-Rottlerin-4

Table 1 MTT reduction normalized to cell numberTime (h)MTT/cell numberR5R2011.231.5421.201.5341.201.70151.252.24241.583.49The MTT (% control)/cell num

Critical-Appraisal-of-the-MTT-Assay-in-the-Presence-of-Rottlerin-6

Our experience indicates that it may not be sufficient to change the medium containing Rottlerin and to wash the cells before adding MTT to avoid a po

Critical-Appraisal-of-the-MTT-Assay-in-the-Presence-of-Rottlerin-5

Because an immediate sign of mitochondrial uncoupling is the drop in cell phosphorylation potential, we evaluated the Rottlerin uncoupling effect by d

MTT-Assay

 This procedure is for cells in 96 well plates, if larger plates are used then adjust volumes accordingly.1 Make a solution of 5mg/ml MTT dissolved in

MTT-Cell-Proliferation-Assay

MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay, first described by Mosmann in 1983, is based on the ability of a mitochondri

Cytokine-Bioassays

Cytokine BioassaysIntroductionBiological activity of cytokines and their concentrations are commonly measured by cellular proliferation of primary cel

Neutralizing-Bioassay-Protocols

Neutralizing Bioassay ProtocolsIntroductionAntibodies that block binding of cytokines to their specific receptors and neutralize their effects are cri

Hanging-drop-aggregation-assay

DescriptionThis assay is used for the aggregation property of cancer cells. It is a very critical parameter for measurement of cell metastasis. Factor

TCell-Activation-Using-mAb-to-CD3

IntroductionMature T cells recognize and respond to the antigen/MHC complex through their antigen-specific receptors (TCR). The most immediate consequ

Culture-of-rheumatoid-arthritis-synoviocytes-and-collection-of-synovial-...

Culture of rheumatoid arthritis synoviocytes and collection of synovial fluidsCulture of rheumatoid arthritis synoviocytes and collection of synovial

Aspartate-Assay

实验概要The  Aspartate Assay Kit provides a simple, convenient assay to measure  aspartate in a variety of samples. In the assay, aspartate is converted  

Phosphate-Assay

1. Make standards using sodium phosphate at the following uM concentrations: 0, 2, 5, 7, 10, 20, 40, 60, and 80. Use the screw top glass tubes.2. Dry

Protease-assay

实验概要        In certain fruits, such as pineapples and mangoes, the flesh contains protein-digesting enzymes (proteases). These may play a part in

TUNEL-assay

PROTOCOL:•Deparaffinize and rehydrate slides:3 x 3´ Xylene3 x 2´ 100% ethanol1 x 2´ 95%, 80%, 70% ethanol (each)1 x 5´ 1x PBS•Microwave antigen retrie

Protease-assay

In certain fruits, such as pineapples and mangoes, the flesh contains protein-digesting enzymes (proteases). These may play a part in helping to softe

Bradford-Assay

Bradford AssayThe bradford dye-binding assay is a colorimetric assay for measuring total protein concentration. It involves the binding of Coomassie B

Polygalacturonase-assay

This enzyme is famous for being involved in the development of the GMO tomatoes (more information from the link at the foot of this page). The cells o

Motility-Assay

DescriptionVarious phenotypic characteristics are requiredfor a cancer cell to successfully complete the metastaticcascade. Among these, acquisition o

Bradford-Assay

The bradford dye-binding assay is a colorimetric assay for measuring total protein concentration. It involves the binding of Coomassie Brilliant blue

Pectinase-assay

Pectinases are actually a mixture of enzymes, which, along with others such as cellulase, are widely used in the fruit juice industry where they are w

DGK-Assay

Buffers:- 2X buffer10 ml 0.5 M imidazol, pH 6.60.21 g LiCl1.25 ml 1 M MgCl21.0 ml 0.1 M EGTA, pH 6.6--> Bring volume up to 50 ml with distilled water.

Chemotaxis-Assay

PurposeThe purpose of a chemotaxis assay is to determine whether your protein or small molecule of interest has chemotactic activity on a specific cel

四唑盐(MTT)比色实验——MTT法

实验方法原理活细胞的线粒体中的琥珀脱氢酶能使外源性的MTT还原为难溶性的蓝紫色结晶物并沉积在细胞中,而死细胞无此功能。二甲基亚砜能溶解细胞中的紫色结晶物,用酶联免疫检测仪在490  nm 波长处测定其光吸收值,可间接反映细胞数量。在一定细胞数范围内,MTT结晶物形成的量与细胞数成正比。 实验材料细胞

凝胶延长分析二型脂肪酸合成的方法

Gel-elongation assay for type II fatty acid synthesisSrinivas KodaliAndrew GalgociSheo Singh Dr.Jun Wang Dr., jun_wang2@merck.com, Merck Research Labo

信号传导

Cytokine Bioassays (eBioscience)Biological activity of cytokines and their concentrations are commonly measured by cellular proliferation of primary c

信号传导

Cytokine Bioassays (eBioscience)Biological activity of cytokines and their concentrations are commonly measured by cellular proliferation of primary c

小鼠脾脏T细胞和人PBMCs的体外T细胞活化实验(二)

材料1X 无菌 PBSAnti-human CD3:Clone OKT3 (Functional Grade, eBioscience Cat. No. 16-0037) or Clone HIT3a (Functional Grade, eBioscience Cat. No. 16-0039)R