Please suggest me an inducer of PPAR alpha protein expression?

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Can someone tell me estradiol inducible gene in brain endothelial cell lines?

I want to know a gene strongly altered by 17 beta estradiol in human brain endothelial cells, so that i can take that gene as a control for the system.

04 May 2017 2,380 3 View

Could anyone guide me on procedure to dissolve and make dilutions of 17 beta estradiol?

I am trying to perform gene expression experiment with 17 beta estradiol using HepG2 Cell lines. Somehow i am not able to get any induction or akteration. It looks like i am doing something wrong...

31 December 2016 7,229 5 View

For how many days i have to culture cells in phenol red free media before giving E2 treatment?

I am using HepG2 cells as the model system

31 December 2016 9,479 1 View

Can someone suggest E2-inducible gene in HepG2 cells as an internal control?

I am using HepG2 cells as the cell model. I am giving treatment of estradiol. But i want to be sure that my estradiol drug is dissolved correctly. so anyone can suggest me a known E2 inducible...

31 December 2016 2,202 0 View

How to decide time points of RNA extraction in microarray experiment?

Hello Experts, How do we decide time points of RNA extraction after giving drug to the cell lines for checking the gene expression profile in a microarray?

31 December 2015 5,992 6 View

Can I use the half life of the drug as the time point?

I don't know the time since all individual genes will be expressed at different time points. Can I use half life of the drug as the time point?

31 December 2015 6,128 1 View

Have someone successfully used the ER alpha (Santa Cruz sc-8002) antibody in immunostaining ?

Santa Cruz now recommends this new antibody (F-10) but I can't find a publication using this reference. I'm working on human et non human primate hypothalamus and so far, it doesn't work. Is...

02 March 2021 10,003 2 View

Energy minimization in LAMMPS using NVT ?

How do I do energy minimization in lammps with NVT ensemble ? I am using the following command and it does not seem to work. It stops at extremely high energy and gives me the error zero search...

01 March 2021 2,294 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

How to replicate the well conditions of a crystal hit with 20%(w/v) isopropanol?

I had a crystal hit in a well with 0.1M HEPEs pH 7.5, 10%(w/v) PEG 4000, and 20%(w/v) isopropanol. Why is isopropanol in w/v here if it's just solvent? How do I recreate this well condition?

25 February 2021 9,761 1 View

Problem with ligating 2.8 kb insert into 8.5 kb plasmid?

I am trying to clone 2 copies of the same mammalian gene into the pSF-CMV-Ub-Puro Ascl (contains HindIII, KpnI and NheI cut sites) plasmid. This is what the final product is supposed to look like...

24 February 2021 6,310 3 View