What concentration do you use to elute a NiNTA resin?

The protocol of Thermo Scientific NiNTA resin (cat. 88221) for purification of proteins containing a hexahistidine tag use an elution buffer containing 250 mM imidazole. For fun after this elution...

07 August 2019 6,108 6 View

Dehydrogenase enzyme activity: would you take a look at my data?

My experiment involve a dehydrogenase and I would like to identify the optimal substrate. For this an assay was established based on the use of DCPIP as cofactor, which when reduced the absorbance...

31 December 2018 9,454 7 View

Potassium cyanide in redox enzyme characterization?

My goal is to characterize a dehydrogenase, we are unsure of which cofactor it can uses but we tried DCPIP as artificial redox cofactor and there is no activity. I would like to try phenazine...

05 June 2018 7,187 4 View

Methods to stop enzyme reactions?

I checked the previous answers :) So my enzyme kinetic assay is based on DCPIP absorbance at 3 mM, which is outside of the linear optical range to measure continuously on a plate reader. Then I...

03 April 2018 8,655 9 View

Why NIST charges for their spectral libraries?

NIST, the National Institute of Standards and Technology publishes reference libraries of hundred of thousand of compounds. I would like to use these in my computer for GC data, however it looks...

03 April 2018 6,523 3 View

What is the CKBB buffer?

In Gao, et al (NAD-Independent L-Lactate Dehydrogenase Required for L-Lactate Utilization in Pseudomonas stutzeri A1501, doi:10.1128/JB.00017-15) CKBB buffer is employed for the optimum pH and...

31 December 2017 3,938 3 View

How high can I go with maltose on a MBP purification?

I have a maltose binding fusion protein being purified with the MBPTrap column (GE). I get very little protein (less than 1 mg from a 400 mL E. coli BL21 (DE3) culture) during the elution step...

10 November 2017 5,667 8 View

Volatile buffers in FPLC: advantages?

What are the advantages and disadvantages of using a volatile or a non-volatile buffer for anion exchange chromatography in an FPLC? Is it only related to further processing , like freeze drying...

10 November 2017 5,422 3 View

FPLC default wavelenghts: what can you measure at 215, 255 and 495 nm?

The default settings of the FPLC UV detector are for wavelengths 215, 255, 280 and 495 nm. I know that the 280 nm is the most important since it directly measures protein via the aromatic ring in...

07 August 2017 9,234 4 View

How can I obtain legible GI sequences?

02 March 2015 6,978 4 View

Steps to calculate activation energy by coats & redfern method?

I'm a chemistry student and I'm researching the kinetic degradation of mixed plastic waste. I'm having trouble calculating the activation energy by the coats & Redfern method. The data that...

24 February 2021 974 1 View

How to calculate the fraction metabolized of a drug by an enzyme using clearance(ml/min) values of the metabolite formed?

I am trying to find the fraction metabolized of a drug by CYP and UGT enzymes. I am unable to get the values of fraction metabolized (Fm) directly from the literature. Can we calculate the Fm of a...

21 February 2021 5,379 2 View

Onset of Photo Oxidation in Polymers?

Hi! I am looking for some specific information in regards to the minimum amount of UV intensity required to begin the photo degradation process in polymers. I understand each polymer is...

15 February 2021 7,703 4 View

How long does it take to design and validate transition state analogue inhibitors?

Hi, I'm asking from the point of view of designing a research plan: How long (approximately) can it take to determine the transition state structure of an enzyme substrate using kinetic isotope...

14 February 2021 2,610 1 View

Kinetics of Enzyme production in Recombinant E Coli?

Hello, I am trying to find kinetic information (such as rate of enzyme production etc.) on the expression of taq polymerase in E-Coli after inserting a recombinant plasmid. Any help would be...

12 February 2021 1,386 3 View

How to match the XRD of a compound for which there is no JCPDS file ? Is there any other characterization to claim the synthesis of desired compound?

I have synthesized LiMgF3 using the starting precursors LiF and MgF2 via solid state diffusion method. Now I am looking for some characterization to claim the desired product, as there is no...

09 February 2021 921 3 View

How can I quantify the color of a solution?

I know colorimetry has been the standard way by way of measuring the concentration of a particular compound (solute) in a colored solution (solvent or measure quantities of a particular compound...

08 February 2021 2,673 1 View

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Hello! I want to determine the activity of my in house Reverse Transcriptase enzyme. For this, I have used Thermo's Enzcheck Reverse Transcriptase assay kit. I used commercial Thermo RT enzyme to...

08 February 2021 7,285 1 View

How define one baker unit(BU) is the amount of hydrogen peroxide catalase ?

07 February 2021 2,006 1 View

Does DNA A-tailing survive after agarose gel electrophoresis and purification?

Hi everyone, I need to clone in pGEM-T vector the result of amplifying a metagenomic library by PCR with a high fidelity polymerase but I only want to clone the fragments of an specific size...

04 February 2021 6,051 8 View