Volatile buffers in FPLC: advantages?

More Luis Peña's questions See All

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 6,221 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 9,556 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 7,325 4 View

Steady state kinetics: time frame between replicates must be the same?

I am measuring dehydrogenases enzyme kinetic parameters via Michaelis Mendel fit, the enzyme activity is determined by the inverse of the slope in the absorbance of a redox indicator when this...

04 May 2018 7,050 0 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 8,823 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 6,627 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 4,025 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 5,759 8 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 9,940 4 View

How can I obtain legible GI sequences?

I need to retrieve a large list of protein and their related nucleotide sequences. However, batch entrez removes old GI numbers, which in my case are more than half. How can I override this, or...

02 March 2015 7,078 4 View

Which type of compound does lamda max of 218 indicate in a uv-vis spectrum of a partially purified compound through column and TLC?

A crude extract of fungal culture using EtOH was subjected to column and TLC and partially purified compound was obtained. UV vis spectrum of the compound/s has max absorbance at 218nm. The...

11 August 2024 9,801 2 View

Can you connect an HPLC to a Mass Spec only at a certain time point?

Can anyone explain this method? Especially the last statement where it says only at 1.5 to 2.5mins was the MS/MS connected to the UPLC. How is that possible, is it a feature in this specific...

11 August 2024 8,141 3 View

GC-MS retention index prediticon?

Hello experts, Does anyone know any free software about retention index prediction ?

08 August 2024 7,403 2 View

Can I use a HisTRAP column for affinity chromatography?

I'm working on selecting antibodies against a recombinant protein that has a His-tag. My idea is to first bind the recombinant protein to a HisTRAP column and then use this column for an affinity...

07 August 2024 505 3 View

AMAXA Lonza nucleofection lower volume of buffer?

Does anyone tried to do nucleofection with AMAXA by Lonza with lower than recommended amount of buffer in the cuvettes (100 ul)?

07 August 2024 4,616 0 View

Why I can't see any band in SDS-PAGE?

Currently, when I run SDS-PAGE, I don't see any bands at all, even though I used the same material just a day ago and it worked fine.... In our lab, we dilute the 10X running buffer to 1X and...

06 August 2024 5,373 2 View

Why does my protein refolded to beta sheet during thermal denaturation analysis?

Hi! So i attempted to understand a novel protein behavior towards heat application by analyzing its secondary structure change. I subjected the protein to a thermal denaturation analysis using...

06 August 2024 1,989 3 View

Why is electronic energy 0.000 for liquid crystal compounds and is invariable with temperature in Guassian 09 software?

Are there any suggestions or insights you can provide to help address this problem

06 August 2024 1,443 2 View

Why might the impedance values for DI water and 0.1X PBS buffer solution exhibit a decreasing and increasing trend, respectively over time (HP 4194A)?

Hello everyone, I'm encountering an issue with my electrochemical impedance spectroscopy (EIS) measurements and would appreciate some insights. Experimental Setup: Electrodes: Gold interdigitated...

05 August 2024 3,783 2 View

Why is TLC not a form of partition chromatography?

Why is TLC not a form of partition chromatography and paper chromatography not a form of absorption chromatography?

03 August 2024 7,035 3 View

Omar Gonzalez-Ortega

Yes, as long as the buffer is selected appropriately to actually buffer.

Dominique Liger

Hi there,

You can get rid of it just by evaporation without any usual desalting step. Volatile buffer doesn't basically exhibit any inconvenience.

Sara Fernández-Castillejo

Which is the best volatile buffer to lyse cells?