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

Does Napabucasin soluble in dichloromethane and in ethanol?

I need to prepare a solution of Napabucasin (inhibitor of cancer cell stemness) in dichloromethane. The drug is expensive, so maybe somebody already tried to do it. If it is soluble, what is the...

02 March 2021 9,945 3 View

Help request in CST microwave?

im working now on TPV solar cells and metamaterials and i realy need our help in CST microwave i have a tpv cell simulation and i want to extract particullarly the absorbtion of only the active...

02 March 2021 2,246 2 View

Dapi fluorescence protocol for colon cell lines ?

28 February 2021 2,534 2 View

Does RNAiMAX transfection reagent work in serum free medium?

I diluted siRNA and RNAiMAX in opti-MEM and added to the cells which they were in the growth medium. Is it a right way? or should I culture cells in the opti-MEM medium for a while and not in...

26 February 2021 10,041 3 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

Unknown debris sheets observed in cell culture flasks?

Hi, We culture neural progenitor cells in T175 culture flasks coated with PLO-Laminin. We have successfully grown these cells with our protocol for months. Recently, we observed large sheets...

25 February 2021 3,443 4 View

How do I transfect a large plasmid into PC12 cells with nerve growth factor (NGF) receptor?

I have a large 16 kb plasmid, which I need to transfect into PC12 cells. Lipofectamin 2000 didn't work and with GFP alone the transfection rate is very low. I also tried the neon invitrogen...

25 February 2021 6,635 5 View

What is a good reagent for lentiviral plasmid transfection in HeLa cells?

I have been using Lipofectamine 2000 but was hoping there may be a better protocol or one for something like Lipofectamine PLUS reagent.

24 February 2021 1,633 4 View

Is it possible that Cancer cells could survive in heparinized blood for 48 hours?

Hi everyone, I am just wondering if we incubated 200 microliter of whole blood with 50 microliter of cancer cells(1bout 5000cells) do you think that cancer would survive for 48 hours?

23 February 2021 3,840 3 View