Electroporation conditions for primary retinal cell cultures (chick embryo)?

We are trying to transfect primary retinal cell cultures (chicken) with a pCAG (Addgene) plasmid but we have very low transfection efficiency (

07 August 2016 9,853 0 View

Calculation dilemma in a GH ELISA kit designed for media and plasma, Can you help us?

07 August 2016 3,105 6 View

In Response Surface Methodology is it mandatory to always start experimenting with first degree equation? Can one start with a second order one?

02 March 2016 3,798 2 View

What's the best way to shear DNA in a nuclear solution?

I've extracted fractioned protein lysates from adherent breast cancer cells. I've had good results with the cytosolic and mitochondrial fractions. However, the last fractioned obtained is a...

02 March 2016 5,078 4 View

Advice on antibodies for western blot detection of BNIP3/BNIP3L?

Hi, I'm looking into regulation of mitochondrial apoptosis by a novel compound. Gene expression results suggest it induces upregulation of both BNIP3 and BNIP3L and I'm considering looking at...

02 March 2016 2,126 1 View

Has anyone had any experience using the abcam Cell Fractionation Kit (ab109719)?

Hi. I'm interested in using this kit to obtain cytosolic and mitochondrial protein fractions from adherent breast cancer cells and was wondering if anyone had any experience (good or bad!) or...

01 February 2016 1,325 0 View

What's a suitable positive control for detection of PARP cleavage using western blotting?

I have good results showing the induction of PARP cleavage by different agents, but I need to include a positive control for publications. Any suggestions? Thanks!

31 December 2014 5,135 5 View

Basic electrochemistry question: how does the midpoint potential of an analyte affect its redox reactivity?

I've been involved in some analytical electrochemistry experiments to determine the reduction/redox potential of some compounds I'm working with. Cyclic voltammetry of two different analytes has...

10 November 2014 1,270 6 View

Did you collected centipedes?

I could collaborate by identifying scolopendromorph centipedes (Chilopoda: Scolopendromorpha).

01 January 1970 8,970 2 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

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

How to make fractions of chloroform extract of plant, using seperating funnel?

28 February 2021 8,466 2 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

What's the concentration recommend for recombinant protein use in western blotting?

I'm currently working having problem with the concentration for recombinant protein use in western blot, the one been used is Recombinant Human CD19 Fc Chimera Protein-rndsystem which does not...

24 February 2021 8,514 3 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