Search found 8 matches

by dalin
Mon Nov 05, 2012 8:10 am
Forum: General Discussion
Topic: FLU/time to FLU/mg/min
Replies: 6
Views: 5469

Re: FLU/time to FLU/mg/min

Just to also add to this problem! The concentration in mM is supposed to be obtained by comparing the enzyme data with the standard curve in such a way that I convert the RFU (for the enzyme data) into mM using the free Fluorescent reference curve ( consist of mM concentrations of the Fluorescent co...
by dalin
Mon Nov 05, 2012 12:33 am
Forum: General Discussion
Topic: FLU/time to FLU/mg/min
Replies: 6
Views: 5469

Dear Jack, Thanks for your reply. But can you give details how to convert concentration for instance in M, mM, uM into moles, mmoles, nmoles as i am kind of "stupid" sometime with the conversion between concentration and other formula. Please detailed example if you like? also, the n is mo...
by dalin
Sun Nov 04, 2012 7:38 am
Forum: General Discussion
Topic: FLU/time to FLU/mg/min
Replies: 6
Views: 5469

Thanks for your respond. But how I convert RFU that is cleaved from the substrate into uM or mmole using a standard curve with the free fluorescent as a standard reference. I am kind of stuck here how to work it out using excel. Is it right if I plot serial concentrations of my reference fluorescent...
by dalin
Sat Nov 03, 2012 10:38 am
Forum: General Discussion
Topic: FLU/time to FLU/mg/min
Replies: 6
Views: 5469

FLU/time to FLU/mg/min

With the fluorometric enzyme Assay how to convert the FLU/min (obtained as FlU/min slope for every concentration of substrate; serial dilutions of substrate at concentrations 200 uM to 50 uM), into to RFU/mg/min of enzyme, that is nessesary to obtain the rate of reaction for the enzyme. Enz. Eny use...
by dalin
Sun Oct 21, 2012 11:18 am
Forum: Molecular Biology
Topic: Multiple reactions buffer and enzymatic special activity
Replies: 7
Views: 5498

Re: Multiple reactions buffer and enzymatic special activity

Thanks for this explanation, but I still need your help! How to prepare multible reagent buffer how do I calculate and prepare the concentrated buffer that is 10-times higher concentrations of 250 mM Tris, pH 8.0, 1.5 M NaCl, 30 mM KCl, 10 mM MgCl2 and 20 mg/ml BSA. Let's say my final volume is 500 ...
by dalin
Sun Oct 21, 2012 5:29 am
Forum: Molecular Biology
Topic: Multiple reactions buffer and enzymatic special activity
Replies: 7
Views: 5498

In the scientific papers or in protocol procedure, sometime do not understand the way they write their experimental data. For instance, a Buffer that consists of a multible reagents and labeled (10X). I was asked to dilute it by taking 3 ml of that 10x buffer and add 30 ml of distelled water and I w...
by dalin
Wed Oct 17, 2012 3:45 am
Forum: Molecular Biology
Topic: Multiple reactions buffer and enzymatic special activity
Replies: 7
Views: 5498

sorry, i mean U/ml not mU/ml , also U/ug and one unit = 1 pmol/min those refer to how enzyme activity is defined and are important for the design of the conditions and concentrations of enzyme/buffer/substrate/co-substrate. Please help me?
by dalin
Wed Oct 17, 2012 3:40 am
Forum: Molecular Biology
Topic: Multiple reactions buffer and enzymatic special activity
Replies: 7
Views: 5498

Multiple reactions buffer and enzymatic special activity

In the scientific papers, If get stuck with the way they write that is kind of arbitrary for instance if you have an assay Buffer (10X) that is concentrated and if I dilute 3 ml of that by adding 30 ml of water which will bring the buffer concentration to 25 mM Tris-HCl, Ph 8.0, containing 140 mM Na...