| | SLO | ENG | Cookies and privacy

Bigger font | Smaller font

Show document

Title:Optimizacija metode za ocenjevanje antioksidativne aktivnosti z uporabo barvila dcfh-da na celični liniji tlt
Authors:Oswald, Barbara (Author)
Langerholc, Tomaž (Mentor) More about this mentor... New window
Files:.pdf MAG_Oswald_Barbara_2016.pdf (1,39 MB)
MD5: 1ABE58EBE77D941AC32493E61F898731
 
Language:Slovenian
Work type:Master's thesis/paper (mb22)
Organization:FKBV - Faculty of Agriculture and Life Sciences
Abstract:Na modelni makrofagni človeški celični liniji TLT smo optimizirali celično metodo z uporabo barvila DCFH-DA (2´, 7´-diklorofluorescin-diacetat) za določevanje pro- in antioksidativne aktivnosti modelnih substanc vitamina C in troloksa. Ugotavljali smo najprimernejše število celic na vodnjak, optimalen inkubacijski čas in koncentracijo testnih substanc ter najprimernejši vrstni red nanosa reagentov. Vrednost fluorescence v testu z DCFH-DA je bila linearno odvisna od števila celic na vodnjak, vendar le v odsotnosti oksidanta t-BuOOH. Pri eksperimentih se lahko uporablja širok interval koncentracije celic, saj so vse vrednosti izmerjene fluorescence še vedno bile v območju detekcije inštrumenta. Optimalen inkubacijski čas troloksa je od 1 do 2 uri, vitamina C pa 6 ur ali več. Visoke koncentracije obeh antioksidantov lahko vplivajo na nastalo fluorescenco, če sta prisotna v fazi merjenja. Optimalna vrstna reda nanosa reagentov sta bila testna substanca - DCFH-DA - t-BuOOH in t-BuOOH - testna substanca – DCFH-DA.
Keywords:antioksidativna aktivnost, DCFH-DA, celice, vitamin C, troloks
Year of publishing:2016
Source:Maribor
NUK URN:URN:SI:UM:DK:PIRNOT6A
Views:1732
Downloads:119
Metadata:XML RDF-CHPDL DC-XML DC-RDF
Categories:FKBV
:
  
Average score:(0 votes)
Your score:Voting is allowed only for logged in users.
Share:AddThis
AddThis uses cookies that require your consent. Edit consent...

Hover the mouse pointer over a document title to show the abstract or click on the title to get all document metadata.

Secondary language

Language:English
Title:Method optimization for tlt cell line based antioxidant activity assay using dcfh_da flurescent probe
Abstract:We have optimized a TLT cell-based method using 2´,7´-dichlorofluorescin-diacetate (DCFH-DA) to assess the pro- and antioxidative activity of model substances vitamin C and trolox. We tried to determine the most appropriate number of cells per well, optimal incubation time and concentration of tested substances and the most appropriate order of reagent application. The fluorescence value in DCFH-DA assay was linearly dependent on the number of cells per well, but only in absence of the oxidant t-BuOOH. A wide range of cell concentrations can be used in this assay, that were within limits of instrument sensitivity. Optimal incubation time for trolox was 1 to 2 hours, while 6 or more hours are required for vitamin C. If those antioxidants are present while the measurement is conducted, their high concentrations can influence the resulted fluorescence. The most appropriate orders of reagent application were tested substance - DCFH-DA - t-BuOOH and t-BuOOH - tested substance - DCFH-DA.
Keywords:antioxidant activity, DCFH-DA, cells, vitamin C, trolox


Comments

Leave comment

You have to log in to leave a comment.

Comments (0)
0 - 0 / 0
 
There are no comments!

Back
Logos of partners University of Maribor University of Ljubljana University of Primorska University of Nova Gorica