How can I measure PKA activity in the yeast cell?

Hi everyone. I would like to measure PKA activity in the yeast cell. Could you give me any suggestions? Thank you!

05 June 2016 3,771 0 View

Does anyone know a protein marker which is only expressed after diauxic shift or in stationary phase in yeast?

Hello everyone, I am going to analyse some protein targets in stationary phase in some mutants. However, the mutants grow slowly, so I want to use some indicative proteins to determine the...

08 September 2015 7,542 2 View

How to dissolve polylysine?

Hello everyone, I wanna attach my cells on to the slide, so I used poly-lysine to coat the slide. I made poly-lysine solution in ddH2O with concentration was 1mg/ml and stored at -20oC. After...

01 February 2015 1,309 6 View

Is it difficult to express small recombinant protein in E. coli?

31 December 2014 6,670 6 View

Should I insert a simple peptide between GFP and target protein?

Dear everyone, I want to tagged GFP to my target protein. Do I have to insert a simple peptide such as GGGG between GFP and the protein? And how many amino acids is enough? Thanks you so much.

11 December 2014 7,104 8 View

Do you know where to get information about the yeast backgrounds?

Hi everyone, I am working with BY4741 background but it seem to be not working well. So I want to change to another background such as CML128,... But I do not know much about the other backgrounds...

11 December 2014 1,414 0 View

I'm making the trypsin 0.25% solution for cell culture. The recipe is 2.5g trypsin and 0.2g EDTA.4Na in 1l HBSS 1x. My lab only has EDTA.2Na. How can I change EDTA.4Na to EDTA.2Na?

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04 May 2013 6,078 9 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

Which is a lucid standard comprehensive book to guide on psychological scale construction and its standardization?

02 March 2021 7,082 3 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 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

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View

What is the most suitable server for predicting protein structures in bacteria?

I have used the i-Tasser several times, however it has been unavailable for several days. I tried the swiss-model, but the output was not very pleasant due to the model used. Is there any other...

28 February 2021 4,521 3 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

Are protein samples in SDS compatible with ELISA?

I have used an AllPrep DNA/RNA/Protein Mini Kit QIAGEN kit to extract RNA and protein from my samples. At the end of the protein purification, I resuspend my protein in 5% SDS. Will these samples...

28 February 2021 7,370 3 View

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View