How to prepare calibration buffers to measure lysosomal pH properly ?

06 June 2018 1 9K Report

Hello,

I would like to measure lysosomal pH using a ratiometric probe, and in order to convert the fluorescence ratio into absolute pH values, I would like to use "calibration buffers".

I saw that nigericin/monensin are commonly used in this goal.

However, I do not succeed in finding a detailed protocol to prepare this kind of buffers. Could someone help me ?

Many thanks,

Yohan

Viorica Raluca Contu

I am trying to do the same and I recently found this paper. It is probably too late but I hope it helps. I haven't figured out yet why adding nigericin/monensin is necessary, though.

Article Live-cell Microscopy and Fluorescence-based Measurement of L...

Badges
Science topic

More Yohan Santin's questions See All

Is it possible to generate a double tamox inducible-Cre mouse?

Hello, I was wondering if it is feasible/easy to induce a double knockout in mice by employing a strategy that involves crossing two mouse strains, each bearing a tamoxifen-inducible Cre (CreERT2)...

23 January 2024 5,848 0 View

Have someone successfully used the ER alpha (Santa Cruz sc-8002) antibody in immunostaining ?

Santa Cruz now recommends this new antibody (F-10) but I can't find a publication using this reference. I'm working on human et non human primate hypothalamus and so far, it doesn't work. Is...

02 March 2021 10,460 2 View

Monitor conformational changes in a protein influenced by pH?

I work with a protein that apparently likes acidic pH and I suspect that it do change its structural conformational when I change the pH. Apart from crystallography, NMR, circular dichroism, gel...

19 March 2020 6,430 3 View

What is the best signal processing approach for EEG signals?

EEG signals consist with lot of noise. Fast Fourier Transform (FFT) , Short-Time Fourier Transform (SFTF) are some of known techniques for pre-process these signal. Is there any better solutions?...

24 May 2019 5,180 4 View

Which data can be taken from EEG signal to analyze lie detection?

EEG signals consist with very noisy data. How those signals use to perform task like lie detecting?

19 April 2019 6,151 1 View

How can i calculate the initial rate of reaction in Fret analyses?

Hi everybody, I performed a Fret analyses of Cathepsin. I add different concentration of fluorescence substrate and i blocked the reaction after 90 minutes. I obtained RFU but now i don't know how...

21 March 2019 8,689 1 View

Can Sepharose-Nanobody-(GFP) columns be regenerated after a GFP-tagged protein purification?

I am going to purify several proteins by using GFP-nanobodies linked to a sepharose column since my proteins have a GFP taq. Since I have not experience with this protocol, I would like to solve...

20 November 2018 1,769 1 View

Does all the Baculovirus Bac-to-Bac-like systems share donor plasmid compatibility?

I am going to clone several protein to express in Baculovirus. I am clonning them using one plasmid that I know is compatible with the Bac-to-Bac system that is being used in the lab, (since is...

25 October 2018 2,024 1 View

What is the best reliable primers set /protocol for SYBR Green I based quantitative detection of Dengue and nested PCR for sero-typing?

Presently I'm using Lanciotti et al., protocol for detection of Dengue using SYBR green1 PCR and I need to extend it for quantification of copy number / viral load in patient serum samples. I...

19 September 2018 8,872 1 View

Interaction effect for L2 acquisition

Would you advise me of the any literature regarding how interaction affects to L2 acquisition? Interaction is like not only classroom interaction but also social interaction such as social...

22 May 2017 334 3 View

How to assess the osmotic power of a substance?

Hello, I suspect that a molecule I am using is causing bacteria to experience osmotic shock. What chemical properties should I look for to compare this capability with those of other substances?...

06 August 2024 977 1 View

How to check pH of a high strength hydrogel?

We are working on biopolymeric hydrogels. Our system is highly viscous and sticky, and the gel formed are high in strength. We are unable to use pH electrode and pH strip. Please suggest an easy...

30 July 2024 942 2 View

EDTA buffer solution?

Hello everyone! How can i prepare EDTA buffer solution (pH 8.2) to determine GSH levels? Thank you in advance.

29 July 2024 4,374 1 View

Whether i can preserve leaves samples in 2.5% glutaraldehyde in 0.05 M phosphate buffer, pH 7 for coming several weeks for further LM study ?

I want to preserve the sample leaves for the LM study of the stomata and conidia analysis. Whether 2.5% glutaraldehyde in 0.05 M phosphate buffer with pH 7 can be applied for preservation.

29 July 2024 8,560 0 View

How can I regenerate a SCX column modified with Sulfonate group (Thermo 054995)? The colmun has lost its Resolution and peak to valley ratio?

The column is used in Ion Exchange Chromatography of Monoclonal antibodies and the method of analysis is based on pH gradint.

13 July 2024 7,462 1 View

My Cyclic voltammetry curve doesn't show cathodic peak of Cu2+ to Cu+. how to solve this problem?

I performed CV of Cu-MOF in a buffer solution of pH 8, and we did not find any peak corresponding to Cu2+ to Cu+. What is the reason behind this? Please give your valuable suggestions.

08 July 2024 2,388 4 View

Is it possible to generate a (semi-)permanent "transcription bubble" within a replicable plasmid?

I was wondering if it is possible to form a permanent open "ssDNA bubble" similar to a transcription bubble (>13 nucleotides) within E. coli. These criteria are important: 1. Open ssDNA...

07 July 2024 8,275 5 View

Has anyone used the new E1 Clip tip digital pipettes?

We just purchased a multichannel and a single channel E1 Clip tip pipettes. In our first attempt to use them for an experiment they are not dispensing accurately? The rep told me that...

01 July 2024 1,983 0 View

Is it possible for the soil pH to be around 8, but with negligible amounts of carbonates, bicarbonates and exchangeable sodium?

Can this pH of 8 be due to the presence of other substances? Or do I have an error somewhere in the analysis? Thank you!

26 June 2024 9,093 4 View

Which pH range is allowed for subcutaneous pump administration?

Hello, We are designing a protocol to determine the effect of certain compounds in the health of rats, but our problems have certain instability issues when subjected to certain pHs. We were...

19 June 2024 5,950 1 View