How could a transcriptional factor purified as inclusion bodies (by using urea) exhibit a ratio 260/280>1?

I am working with a very unstable transcriptional factor. I use DNAse in the lysis buffer and I purify it as inclusion bodies, by using urea and 1M NaCl. After purification, I buffer exchange the...

05 February 2021 2,530 3 View

Is there some good Instrument for evaluate life's quality of patients with chronic disease?

I'm trying to conduct a research from this field. There's someone to help me?

09 December 2020 4,065 5 View

Do you have any suggestions for a pre-treatment of UF of a industrial liquid waste with high concentration of colloids?

We are working on ultra-filtration of a liquid waste but there are some problems with membrane fouling. Do you have any suggestions of suitable pre-treatments for a sample with high concentration...

19 November 2020 2,122 6 View

How to get higher concentration of linearised plasmid?

I am Trying to use Infusion cloning to prepare a mutant construct. I have been told by a student that used it before that I need 20ng/ul of linearised plasmid to proceed with the following...

04 April 2019 646 10 View

How to remove RNaprotect from lysited cells before RNA extraction?

04 February 2016 1,411 9 View

How can I get the retina remains stuck in its place during histological process?

Eye fixation for histology

15 September 2014 2,371 9 View

Wortmannin increases the levels of LC3-II. Any idea about why this is happening?

I am using wortmannin as an inhibitor of autophagy. When I check autophagy inhibition by WB of LC3-II, in some cell lines wortmannin treated cells have more LC3-II than untreated, contrarily to...

27 April 2014 8,669 4 View

What is the best way to analyse a three-way choice experiment?

I am looking to analyse a three-way choice experiment but am struggling with the non-independence of the data. The data is generated by animals choosing to spend time in three environments within...

04 March 2014 3,778 7 View

Is anyone working on designing sustainability indicators or sustainable development goals that reflect the values of indigenous communities?

Global discourses of sustainability and post-2015 Sustainable Development Goals tend to be based on the Brundtland definition (meeting the needs of the present without compromising the ability of...

12 December 2013 316 17 View

Does anyone have experience using pSTAT 1 and 3 antibodies for western blots?

I have been struggling to detect pSTAT 1 and pSTAT3 bands on my western blots. I am running 10% SDS-PAGE gels for approx 90 mins at 100V, transferring at 80V for 90 mins and blocking in BSA. I am...

29 August 2013 6,408 15 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

No title

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

No title

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