What's the max. passage on HSKM cells / can they be grown/split/differentiated similarly to C2C12 cells?

We are considering a move from C2C12 cells to HSKM cells since the former is not applicable for GLUT4 studies, however we don't have any experience with human cell lines. My questions are:1. What...

13 May 2016 4,306 1 View

How can I best determine extracellular (media) glucose concentration of differentiated C2C12 cells (non-radiometric)?

I'm trying to measure the extracellular glucose of my cells to get a feel for their rate of glucose uptake (from a fixed concentration of glucose), however I'm not really an experimentalist and am...

10 January 2016 9,445 3 View

Can the same sample with a negative result in conventional PCR, give a positive result with real time PCR?

14 October 2014 364 5 View

Does anyone know what genus and/or species of Cladocera this is? Or if it is Cladocera?

I found it with a phytoplankton net in the Oslo fjord, right near the shore. It's quite small compared to most of the Daphnia I've seen. About the size of a large (or two tiny) Ceratium longipes...

11 August 2014 4,361 19 View

QPCR: reference gene stable under hypoxic conditions?

I'm trying to compare mRNA expression levels of my gene of interest in cells at normoxic vs. hypoxic (i.e. 1% O2) conditions. Usually I would use GAPDH as my reference gene but it seems that GAPDH...

31 March 2014 5,003 48 View

Identified vs anonymous survey answers

Hi. When delicate topics like perceived social norms regarding immigration, violence, racism etc are measured and political affiliation needs to be considered, what is the opinion on identified...

01 January 1970 8,181 4 View

Can anybody recommend what measure might predict how open children are too difference?

The study is based of first impressions of autistic children.

01 January 1970 7,233 1 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 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

What urea buffer can i use for biotagged protein extraction from streptavidin beads?

Hi all, I have been doing research on biotagged LDB1 proteins in Mel cells. I purify the proteins using M280 streptavidin beads. The yield is very low so I've been trying to optimize it by...

24 February 2021 5,749 3 View