Suitable sample types for assaying cellulase activity?

Hello, I am trying to set up an enzymatic assay to determine the activity of cellulase. I would like to know what sample types would be suitable to perform the same? I have read certain papers...

01 February 2020 1,105 3 View

SEC for exosome isolation?

Hello, I am trying to isolate exosomes from serum using SEC protocol. I have observed that the fractions mostly collected during the process are very diluted making it hard for me to quantify the...

08 September 2019 5,633 2 View

Positive control for exosome isolation from serum?

Hello all, I am trying to isolate exosomes from serum and would like to know if there is some means to use the whole serum as a positive control? In my previous lab, I used to use whole-cell...

08 September 2019 5,582 6 View

Which running buffer is more compatible to detect TSG101 for exosomes?

I am trying to standardize exosome isolation protocol and want to run a western blot to detect TSG101. Currently, our company does not have a Tris-glycine based system and have been using MOPS...

06 July 2019 969 2 View

Exosome miRNA sequencing core facilities?

Hello, I have exosomal RNAs isolated from serum and plasma specimens of healthy and patient cases whose concentration is very low. The core facility at our university requires at least 1-10 ng to...

03 April 2019 8,504 3 View

Does ETOH-NaOAc precipitation on RNA samples isolated from exosomes increase the ribosomal RNA content?

I precipitated one of my exosome samples using the ethanol-sodium acetate process since my concentration of total RNA was pretty low. Initially, when I checked my RNA content in my exosome sample...

04 May 2018 3,805 1 View

Can we concentrate the RNA that was eluted in elution buffer?

Hello, I performed the total RNA isolation on my samples using miRCURY cell and plant kit, that also helps in preserving miRNAs. The last step of elution, asks me to elute the sample in 50 ul of...

03 April 2018 3,047 8 View

Did someone use the "High Select Top 14 Abundant Protein Depletion Mini Spin Columns" for depletion of abundant proteins and were successful?

Hello, I plan on using the depletion kit on my exosome pellets, since I only see a lot of albumin and other serum proteins. Has anyone used this kit and attained success in getting rid of the...

03 April 2018 7,632 0 View

How to increase the concentration of RNA?

Hello, I tried isolating total RNA that preserves miRNA from 1.2 ml of patient DLBCL serum using miRCURY kit and obtained a concentration of just 9.7 ng/ul on nanodrop. I plan on using this...

31 December 2017 9,060 4 View

Why is the poly A tailing and ligation step necessary in Taqman Advaced miRNA kit?

Hello, I know that poly A tailing and ligation are the two additional steps used in the Taqman Advanced kit in comparison to the traditional Taqman miRNA kit. Can someone explain what is the...

31 December 2017 5,297 7 View

Can we classify HV and LV lines based on line length?

I have dataset which shows the length of power lines. I need to classify the lines based on the line length. Is there a rule to classify the High voltage (HV) and low voltage (LV) lines based on...

03 March 2021 4,116 4 View

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02 March 2021 3,060 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

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

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

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28 February 2021 9,936 2 View

Has anyone had luck using cell lines other than HEK 293 (such as COS1) to package lentivirus?

I am looking to package lentivirus in endothelial cells but I am opposed to using 293 cells due to personal ethics regarding the source. I have done some reading and saw that COS1, COS7, & CHO...

25 February 2021 3,030 1 View