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

What can I use to dilute protein for gelatin zymography?

My protein sample is extracted with RIPA + PMSF ( Phenylmethylsulfonyl fluoride). After protein assay, the concentration is too high (>900ug/mL) for direct application for gelatin zymography...

13 December 2020 7,764 3 View

Why my samples stuck in the top of gelatin zymography?

I am trying to detect MMP2 activity through gelatin zymography. I do can see clear bands. But all my bands stuck in the top of gel. however the ladder passed through...also when i run the gel, the...

03 December 2020 310 2 View

Best choice for Casein Zymography?

I am currently working on a project focused on MMP3. I need to perform casein zymography at some point, but I can't find a proper precast gel containing casein since the product is "discontinued"...

20 September 2020 7,772 3 View

What should be a loading control in condition media for zymography?

I want to do a zymography in condition media from cancer cell line...I would like to know what should be a loading control for condition media specific for zymography? Whether loading control can...

13 November 2019 1,697 1 View

What should be a loading control in condition media for zymography?

I want to do a zymography in condition media from cancer cell line...I would like to know what should be a loading control for condition media specific for zymography? Whether loading control can...

10 November 2019 3,565 1 View

Can mercaptoethanol speed up protein migration in SDS-PAGE?

We have done zymography and Western blot from the same samples. In zymography the detected band is at the size of 90kDa, but in Western blot at 70kDa. At both methods samples are in SDS-loading...

09 October 2019 1,589 7 View

Can I do zymography but the enzyme is mix with acrylamide and the protein is running on the gel?

I want to characterize protein with zymography but the enzyme mix with acrylamide while the protein running on the gel. Is it possible to do that?

08 August 2019 2,918 3 View

Can I do zymography but the enzyme is mix with acrylamide and the protein is running on the gel?

I want to characterize protein with zymography but the enzyme mix with acrylamide while the protein running on the gel. Is it possible to do that?

07 August 2019 3,197 3 View

How much protein from brain extract for gelatin zymography?

Hi! I would like to analyze the activity of MMP2 and MMP9 in extracts from hippocampal tissue. I tried to do it several times but I can't get any bands. I followed Abcam's protocol, which I...

07 August 2019 9,543 2 View

No title

18 July 2019 5,164 3 View