Why has my protein transfer using fresh transfer buffer is worse when compared to protein transfer using old buffer?

Hello, I have performed two westerns in parallel, one using fresh buffer (made with old 10X tris-glycine) and the SDS gel was made 4 days prior to western; and the other western was performed...

03 February 2021 9,311 5 View

When sequencing a plasmid to confirm transformation has been successful, would I have to sequence the full plasmid?

I have transformed some DH5alpha cells with point mutated proteins placed in pcDNA plasmid and extracted the plasmid; now I have to send the plasmid for sequencing for confirming the presence of...

19 January 2021 5,842 3 View

How can decrease the fluorescence background for Raman Spectroscopy?

Could you help me with, how can I decrease the fluorescence background for Raman Spectroscopy?

18 May 2020 7,685 11 View

What are the differences between Raman spectroscopy of amorphous and crystalline carbon materials?

What are the differences between Raman spectroscopy of amorphous and crystalline carbon materials?

30 April 2020 3,435 8 View

In ARDL model, coefficient of independent variable not showing significance. Can it happen? How should i read my results?

Hi, i am working on macro economic data in a bi-variate framework with GDP as dependent variable and inflation as independent variable. I have applied bound test plus ARDL model and it confirms...

13 April 2020 3,375 4 View

How to separate the Raman spectroscopy peak?

In the result of the test for Raman spectroscopy, there are many broad peaks that including a lot of information. In fact, How can I correctly separate those peaks?

26 December 2019 3,460 3 View

Any idea about peak identification of non-expandable minerals in EG treated oriented mounts?

During XRD analyses of oriented clay mounts, a number of peaks which are NOT typical of expandable clay minerals have been observed on XRD patterns of EG solvated ones. Ethylene Gycol was provided...

08 January 2019 6,770 1 View

Why there are two protein chains in Glucocorticoid receptor - DNA binding domain?

Hi, I'm doing a MD study about DBD of Glucocorticoid receptor. Why do it have two Zn motifs ( protein chains)? Both have same function or different functions? What is the importance of having two...

29 August 2017 2,548 3 View

How to do a Molecular Dynamics simulation for DNA molecule without constraint it?

how to do a MD simulation for DNA molecule using AMBER force field in GROMACS? And how to make an one itp file for two complementary strands of DNA?

16 August 2017 7,180 5 View

What is the best way for poor Nations to conserve thier water resources especially with the current trends of climate change?

I am developing a research on the link between poverty, natural resources management and climate change in Nigeria and need some guidance.

11 December 2016 7,577 0 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

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

Can someone suggest a way to prepare slides to observe under confocal microscope for observing fluorescent tagged proteins??

I am growing the cells on coverslips and transfecting the fluorescent tagged protein directly on them. Then i want to observe these cells under confocal microscope. Currently i am just using...

01 March 2021 6,142 3 View

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 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