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Questions related from Erome Daniel Hankore
This has really been quite frustrating. Every time I load my samples (20 to 45 ug per well), I see lines form in the resolving gel and they interrupt the run. The proteins just accumulate on that...
12 December 2019 7,487 10 View
I prepared M9 medium by autoclaving the M9 salts and glucose separately. Calcium chloride, magnesium sulfate, and trace minerals were dissolved in autoclaved water and sterilized by filtration....
06 June 2018 4,883 4 View
I am noticing that in thermostability assays call for heating the enzyme at the desired temp for a set time and then immediately placing in ice before carrying out the assay (as reported in...
03 March 2018 8,477 0 View
Any suggestions? Perhaps in NaOH and the neutralize to pH 7. I need the pH to be around 7 in the final solution.
02 February 2018 5,518 8 View
If I am using substrate concentration that is much lower than the reported Km of an enzyme, say for example 30x lower, is there a way to compensate for that? Can I just add an excess of enzyme...
01 January 2018 4,699 3 View
So, I'm doing an enzyme assay and there are two compounds in the assay, one a product and the other a cofactor, that absorb at the same wavelength. It makes it very difficult for me to detect...
01 January 2018 3,585 3 View
What kind of chemical agents can I use to oxidize NADPH in a reaction and stop an enzymatic reaction? Suggestions other than chemical agents are also welcome. Thanks!
01 January 2018 1,742 5 View
When you have an antibiotic resistance gene in a plasmid, how do ensure that the enzyme is constitutively expressed so that your selection always works?
11 November 2017 5,201 5 View
I am doing some enzyme assays with a non-native substrate of an enzyme. At a certain concentration, it seems to plateau (as if reaching Vmax) and the graph resembles MM kinetics but when I try...
08 August 2017 2,175 15 View
[E]: 10 ug/mL Vmax: 0.098 U/mg Protein mw: 128 kDa Reaction volume: 150 uL I'm having trouble getting to S-1 units for Kcat determination. Kcat = Vmax/[E] so I should divide 0.098 U/mg by 10...
06 June 2017 2,497 0 View
I've been thinking a lot about this question of late. I suppose most people determine kinetic parameters in terms of Km and Kcat to see how well their substrate is catalyzed by an enzyme. But if a...
06 June 2017 5,207 11 View
I am trying to make NADPH solution but because of its hygroscopic nature, it has absorbed water and it is hard to mass it out. My current plan is to just dissolve it in 10 mM NaOH and quantify it...
12 December 2016 4,866 3 View
Thanks.
02 February 2016 1,868 0 View
