How to prevent clumping of EBV trasnfected lymphocytes?

I am culturing lymphocytes and they tend form visible clumps.. Does anybody know how to avoid this and if this is common ? I am new in culturing them. Thanks

04 May 2016 9,428 8 View

Could you share a protocol to culture lymphocytes ?

I am going to culture lymphocytes (EBV immortalized) for the first time and I am looking for protocols to help me with it. I don't see a lot of general culturing protocols online. Could you please...

03 April 2016 5,173 4 View

Contamination problems in 293 T cell culture. Any help ?

31 December 2015 1,872 17 View

Which antibody could I use against BRCA2 for Immunofluorescence?

I am looking for an Antibody to use in Immunofluorescence specific for BRCA2. I am not able to find a convincing IF image of BRCA2 by IF. Lot of scientists use the tagged version of BRCA2 in their...

08 September 2015 3,753 3 View

What are the ways to make the protein marker visible in a WB imager?

I have tried WB specific markers, but I would like to somehow image my protein markers while imaging. Is there a pen that would help ? Or are there other innovative ways?

04 May 2015 4,530 3 View

Has anybody experienced protein sticking to the frits of the Superdex 200 column?

I am working on a human helicase protein and when I load them on a SEC column Superdex 200 (GE), I don't get a good recovery. In addition, when I load my protein-DNA complex into the same column...

31 December 2013 1,388 4 View

How can I stabilise a protein-DNA complex in isolation?

I have purified my protein-Holliday junction complex by SEC, but the purified complex is not stable. The protein dissociates from the DNA. I would want the complex to be stable to perform further...

31 December 2013 3,200 3 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

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

Calculating volume of DNA required from DNA concentration?

Hi I am a bit confused. They are asking me to find out the volume of DNA required in ul (a total of 30-100 ng for genomic DNA) from the DNA concentration in the nanodrop reading which was 404.8...

26 February 2021 5,029 2 View

Has anyone used the Illumina ITS1 primer sets?

Hi everyone, Illumina provides a list of primers to amplify with high taxonomic coverage the ITS1 region for further fungal sequencing, but I cannot find the exact amount necessary of each...

25 February 2021 6,969 3 View