EMSA troubleshooting. Protein-DNA complex not migrating, cold probe lane with multiple bands. Can anyone help?

I'm running an EMSA to show the DNA binding activity of recombinant proteins versus the Wild type. Previous assays run by my research group using a different test system show the protein-DNA...

24 February 2021 8,325 1 View

Why is my qPCR suddently not working?

I've been working with qPCR for some months now. I amplify 2 genes (PSEN2 and HN1), and the protocols are optimized for both and have been working fine. The other day I amplified HN1 for some...

21 February 2021 3,437 3 View

Can ethanol be used as an analgesic?

If ethanol has analgesic properties, why isn't it used for clinical purposes?

18 February 2021 8,227 4 View

Acid phenol:chloroform (25:24:1) for RNA extraction instead of (125:24:1)?

Hi everyone! I am about to extract RNA for RNA sequencing assays and my Invitrogen kit/protocol...

14 February 2021 1,755 3 View

Are there any public datasets of consumer complaints about service industries at the European level?

06 January 2021 1,718 2 View

No title

16 December 2020 9,657 5 View

Tools for analysis of interaction between lncRNAs and proteins/miRNAs?

Hi! I would like to know what tools I could use to analyze interactions between a new lncRNA and possible protein/miRNA targets. The biggest problem I had to find such tools is that the RNA we...

20 November 2020 8,331 3 View

I would like to ask some questions about concentrations of Urea 3M and 8M in two experiments?

I would like to ask if anyone knows if concentrations of Urea 3M and 8M can interfere with protein purification at Ni-NTA and Glutathione Sepharose 43 resins. Another question, Urea, at 3M and 8M...

17 November 2020 5,014 3 View

I would like to ask if anyone among you knows what is the ideal concentration of TFA to use at a chemical synthesis of peptides?

During a chemical synthesis of peptides by the F-moc method, the following happened: all the initial steps of the synthesis occurred as planned, but in the cleavage, TFA is used. In the lab, there...

02 November 2020 2,997 8 View

Can I use generalized linear mixed model to "fix" data absence?

I have some data of the diet of an omnivorous fish diet collected over 20 years. This data is divided in two seasons, the dry period (drought) and the wet period (flood), and I want to analyze the...

10 October 2020 6,546 6 View

How to quantitative analize the data of TNBS assay for gelatin?

Estemeed colleagues, I found some issues regarding the quantification of the data for TNBS assay. There are different protocols on how to perform that but it is clear to me the "fil rouge" that...

02 March 2021 2,616 1 View

Should we change media after1st 6hrs after plating HUVEC bought from INVITROGEN?it's not recomended by invitrogen.Will DMSO present in stock matter?

We have HUVEC bought from Invitrogen. Cat no -C-003-5c. while first plating the cells we have bought the protocol suggests not to centrifuge the cell with media to remove the DMSO. Should we...

02 March 2021 3,713 2 View

Bought HUVEC from Invitrgen.need to revive it.expected it is cryofreezed in DMSO.Do I need to remove the DMSO for plating the cell for the 1st time?

After I cryofreeze the cell do I need to remove the DMSO for culturing the cell for next passages .

28 February 2021 7,738 3 View

For gelatin coating for culturing HUVEC.Should the gelatin be made in sterile water or PBS? Do I need to autoclave the gelatin soln after making it ?

I want to use 0.1% gelatin. Should that be made in PBS or sterile water? Should I autoclave the gelatin soln after making it in water/PBS?

28 February 2021 274 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

Quantifying leaking DNA from bacteria - what's the secret?

Hi, all! I need to determine if my treatment causes membrane damage in bacteria (S. aureus and E. cloacae), but I cannot use a fluorescent probe such as PI and similar ones because my compound is...

24 February 2021 2,006 2 View

How can I verify my centrifugal fan design for electric motor cooling?

I have radial blade centrifugal fan design which will be mounted on exhaust side of electric traction motor. The fan will be mounted on same motor shaft inside the housing. How can I verify that...

23 February 2021 2,390 3 View

Help with culturing monolayer of human brain microvascular endothelial cells (hBMEC)?

I've been having a hard time forming an endothelial monolayer with good tight junctions. The person who trained me on the culture of endothelial cells worked mostly with mouse BMEC, and he used...

22 February 2021 8,253 1 View

Are ZnS nanoparticles moisture sensitive?

Hello Everyone, I tried to form ZnS nanoparticles using the hot injection method and was able to form the same (confirmed by XRD). When I centrifuged, after the reaction I got a kind of white...

17 February 2021 1,709 3 View

How to reduce debris with neuron isolation?

Hi there, I'm isolating neurons from PND1 guinea pigs and have noticed a bit of debris in the early days (this is day 4 plated) and a lot of variability between plates. Current protocol is...

14 February 2021 3,692 4 View