Is there a replacement for succinate buffer (pH-5.8) containing 10mM DTT in cellulase zymography?

I am doing a cellulase isolation and purification from Bacillus sp. I have done the plate assay, and have got about 12 positive results from it, one of it is Bacillus subtilis. I want succinate...

02 March 2015 1,546 1 View

Can you please suggest a most reliable protocol for Cellulase zymography?

I am using Bacillus subtilis as a source for cellulase production grown on 0.1% CMC (Carboxy-Methyl-Cellulose) media. And now I want to detect the presence of cellulase using 0.1% congo red stain...

02 March 2015 7,909 7 View

Do I need to add PMSF (Serine protease inhibitor) separately even after adding the protease inhibitor cocktail?

I am isolating proteins from yeasts.

01 February 2015 10,002 12 View

I want to do a structural comparison of yeast and human metallothionein using bioinformatics tools. Can anybody suggest a suitable tool?

I want to analyse whether a human metallothionein standard for HPLC can be used for yeast metallothionein as well.

31 December 2014 7,516 2 View

Can anyone suggest to me a protocol to measure Metallothionein concentrations in yeast cells using HPLC?

I want to analyse the metallothionein expression level due to affect of heavy metals as a standardization protocol so that it can be extrapolated to mammalian cultures.

09 October 2014 3,492 3 View

No title

02 March 2021 3,060 3 View

Is There Any Feasible Method To Test Fluorescent Compounds Other Than Fluorescence Spectrometers ??

Is There Any Feasible Method To Test The Efficiency Of Fluorescent Compounds Other Than UV Spectrometers ? Suggestions Would Be Appreciated !

02 March 2021 5,785 3 View

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 View

How to accurately quantify chitin?

Hi all, I have been working on the quantification of chitin in insect samples and feed for a while but I can notice that the quantification methods (e.g. exposure to HCl, NaOH, and weighing sample...

01 March 2021 1,843 2 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Can someone suggest a way to prepare slides to observe under confocal microscope for observing fluorescent tagged proteins??

I am growing the cells on coverslips and transfecting the fluorescent tagged protein directly on them. Then i want to observe these cells under confocal microscope. Currently i am just using...

01 March 2021 6,142 3 View

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

What could be some reasons why unstimulated monocytes are getting activated?

Im doing PBMC isolation -> CD14+ enrichment using magnetic beads -> stimulation setup. My negative control is just cells in cRPMI but they seem to get activated over and over again.

28 February 2021 7,883 3 View