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

How to compare two groups with only two measurements?

Hiiiii everyone! I have an enquiry on statistical analysis. I was looking for many forum and it's still cannot solve my problem. I want to compare means of two groups of data but only with two...

03 March 2021 8,796 3 View

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03 March 2021 5,360 2 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

Does anyone know an alternative product for cardiac Troponin T antibody for Immunofluorescence (Thermo # MA512960)?

We have been using Thermo Fisher anti-cardiac Troponin T antibody for immunofluorescence staining in FFPE mouse heart sections but it was recently discontinued. The literature shows it to be the...

01 March 2021 4,937 1 View

Interested to stain mice tissue with WGA 488 conjugate, anybody has a protocol to recommend?

Interested to stain mice brain tissue with WGA 488 conjugate, appreciate if anyone willing to offer me some guidance or suggestions! Thank you!

28 February 2021 3,951 2 View

B-mercaptoethanol (BME) protocol?

Hi everybody, I have been running a multiplex IHC which allow me to stain the same tissue with different markers (Primary Ab + Secondary conjugated Ab) (AF487, AF555, AF647). Since I would like to...

27 February 2021 5,028 2 View

Why do my MCFs not form monolayer?

Hello, I am looking for some expertise on culturing MCF7s. I purchased them from ATCC and I have been following all media and culture conditions. Media is EMEM with 10% FBS and 10ug/mL of...

27 February 2021 3,904 2 View

Is there anyone who experienced with cytotoxic effect of Hoescht 33342 dye?

Is Hoechst toxic to cells? In general, it is known as non-toxic. I am working with dsRED MDA-MB-231 cell line. I applied it for my PDMS-based microfluidic platform, for this reason, I incubated...

26 February 2021 9,119 3 View

Why my agarose gel staining sometimes looks not homogenic but like gradient (see the figure)?

I am worried about this overexposure of the upper part of the gel in the picture. Is it possible that this is the result of too much concentration of ethidium bromide? Why is there such a big...

25 February 2021 8,140 3 View

Markers of mitochondrial activity in FFPE tissues?

Hello everybody, Does someone have any experience with assessing/extimating mitochondrial activity on FFPE tissues? Precisely, no TOM20, ATP5B, etc: we are trying to get an approximative idea of...

24 February 2021 4,618 4 View