How can i detect the phenol red in mixture solution?

Hi. I'm trying to detect the phenol red in the mixture solution by HPLC. As you know, It is using for ph indicator. The problem is.. After making phenol red solution in 50 ml conical tube, I...

13 December 2020 5,511 4 View

How to prevent protein gelation?

Hi :D I have a question. We expressed a protein from E. coli and we got purified peptide. But when we concentrate it with centrifuge filtration (like amicon), It became like a gel. So when we...

05 October 2020 3,727 2 View

Bioanalyzer: Additional Peak in 28s Region?

Hi All, I isolated RNA from human cell line using TRIzol reagent and sent isolated RNA for RNA-seq analysis. The company sent me a QC report showing that some of my samples has additional peak at...

13 September 2020 1,334 3 View

How can solve the problem during Gromac calculation?

Hi :D I'm using Gromacs 5.1.5 for MD simulation. For binding energy calculation, I used g_mmpbsa (version 5.1.2) I simulated the protein for 150ns and nojump of my xtc file. Then cut last 50ns for...

25 August 2020 7,589 1 View

How can i purify my protein sample?

Hi :D I'm doing purify a small protein from E. coli. From E. coli extract supernatant, I purified my protein using IMAC purification with LC. I got a single peak at chromatogram and got the...

23 July 2020 2,393 5 View

DNA fragment digestion without a restriction enzyme site?

Can I digest a specific DNA sequence without a known restriction enzyme site in vitro? I am trying to digest a fragment with CGTCAT but not CATCAT. I tried cas9 with a sgRNA containing the target...

20 July 2020 9,320 4 View

How to determine the number of cells before suspending them in 1x binding buffer in Annexin V FITC assay?

I am performing Annexin V FITC assay for adherent cells. According to the protocol described, the treated cells were trypsinised and collected, followed by washing them twice with cold PBS. The...

18 July 2020 8,695 5 View

How can I prevent protein fragmentation in solution?

Hi, all :D I'm doing short protein expression (~17 kDa, pI 9.6) from Bl21de3 with c-term His tag. I purify it by IMAC (or affintiy chromatography). When I do SDS-PAGE and WB with Anti-His Ab, I...

25 May 2020 6,693 6 View

How to deal with chance constraints in Matlab toolbox ga?

I am learning chance constraints. I want to know how to deal with chance constraints with generic algorithm, especially in matlab toolbox ga? Thank you a lot !

15 March 2020 3,472 3 View

Peptide adsorption to filter membrane? or denatured?

Hi:) I'm doing His tagged peptide expression and purification from E.coli. I expressed the peptide and checked it with WB (with Anti-His Ab). For purification, I did IMAC and got an eluate. But it...

12 March 2020 6,399 13 View

Why does blocking with my immunogenic peptide cause nuclear fluorescence in ICC?

Hi all, I am validating a polyclonal antibody raised against human ANT4, through WB and ICC. In most of my controls, the antibody shows very little off-target binding, but when I pre-incubate the...

03 March 2021 5,999 3 View

Is there any test to determine the cardiovascular risk of patients with obesity? from the perspective of obesity?

We found tests to determine cardiovascular risk taking into account lipid profile, blood pressure, age, history of diabetes, other laboratories. But when weight is involved, there is a paradox. I...

02 March 2021 6,036 1 View

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02 March 2021 3,060 3 View

What SPSS analysis should I run with one IV (with two levels) and two DVs?

Hi, I am trying to figure out which test would be most appropriate to run for the following question: Is there a difference in task blood pressure between smokers and non-smokers? There are two...

01 March 2021 8,235 2 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