Has annyone try to reuse EveryBlot Blocking Buffer (Bio-rad) for fluorescence western blotting?

Hello to every one, Our lab has recently switched our western blot technique from chemiluminescence to fluorescence. Therefore, we have to optimize the blocking buffer. We have successfully try...

05 February 2021 7,886 1 View

Does anyone have experience with movement-based kernel density estimation (MKDE)?

Hello, I have mapped the locations of several individuals of a certain species of bird. I have the GPS coordinates, the date and the time data every half hour for several consecutive days and I am...

26 September 2020 3,712 2 View

Do you know how long it takes for a polyprepylene surgical mask to degrade?

I'm a master student in the university of Geneva. I have to find a scientific paper of the degradation of the surgical mask to add it in my master's thesis. I only found non-scientific journals...

03 September 2020 2,940 3 View

Any suggestion of bibliography in a wide sense (papers, guidelines, systematized experiences) about "male" parenting (fathering)?

Any suggestion of bibliography in a wide sense (papers, guidelines, systematized experiences) about "male" parenting (fathering) and in special in the context of children and adolescents with...

23 February 2020 8,306 3 View

Optimization with Pulp vs Pyomo, any experience?

I have been using PuLP for years and I wonder if refactoring to Pyomo will be faster and/or more reliable, any advice? Thanks in advance!

05 February 2020 3,577 2 View

Why is my protein purification not working?

31 December 2019 3,899 4 View

What is the best way to silanize selective barrels to build K+ sensitive electrodes for extracellular recordings?

We are interested in studying K+ dynamics in cortical slices and to do so we will perform extracellular recordings with K+ sensitive electrodes. However, we have got some problems in buildind...

18 May 2016 8,611 1 View

Blastx of cDNA : gap existence and extension penalties, how important are they?

I am blasting cDNA against a protein database and I was wondering whether the default gap parameters, i.e. big penalty for gap existence and small penalty for gap extension, would be the most...

01 October 2015 7,874 1 View

Is there any public code can calculate r-process chemical abundances?

01 January 1970 4,167 1 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

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 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

Expression cloning by using cDNA,do I need to modify the cDNA first?

I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...

28 February 2021 5,440 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

PUC19 as a transfer plasmid for lentiviral delivery?

Hello, Is it possible to use pUC19 as a transfer vector to be packed in using the second generation viral particles packaging system( pMD2.G; psPAX2 plasmids)? As far as I understand it there is...

28 February 2021 4,868 2 View

RIA. How to measure?

When you use RIA, with a control the unknown sample with antibodies, you bare in mind the binding sites of the antibodies. However you still need to measure labelled antigen (radioactive) and not...

28 February 2021 2,133 3 View