What is the best website or software by which I could determine transcription factor(s) that potentially bind to upstream my gene of interest?

I am trying to find a transcription factor(s) that potentially binds to a specific segment of my gene of interest. The segment is a little bit large (about 3kbp). knowing which transcription...

09 October 2019 2,957 2 View

What does it mean when you have less cells# in flow cytometry plot compared to a control group?

Hi! I have run flow cytometry for 3 different cell lines (black, blue, and red) which were stained with red dye (FL2) with a same event number (15,000 events). However, the cell number (count; Y...

09 October 2017 2,961 3 View

Is there any difference regard to a cell behavior after knock out a gene using different plasmids ?

01 February 2017 1,324 3 View

Is there any ideas why the over expressed protein behaves differently than the endogenous protein?

09 October 2016 916 9 View

What is the best way to test several genes expressions using qPCR?

06 July 2016 5,179 3 View

What is a good or official program to quantify western blotting results?

What is a good or official program to quantify western blotting results? Any help would be appreciated and thanks

08 September 2015 1,600 2 View

Does someone know a web sit(s) by which we can check the biological conferences in the USA?

03 April 2015 909 0 View

Is there any way to avoid cleavage of eukaryote protein expressed in E coli?

Hello, I am trying to express eukaryote protein (36 KDa) in E coli ( BL21 and Rosetta) by using different vectors (pGEX-4T-2 and pET-28a) and tags (GST and His- tag, respectively). However, more...

03 April 2015 7,830 8 View

How many times can we use GE Healthcare GSTrap FF, 1 ml?

31 December 2014 4,600 0 View

Is there any way to purified a protein that does not bind in either cation column nor anion column?

Hello, I express my protein with GST tag and trying to get my protein pure without the tag. Therefore, after I got GST-protein partially pure by performing affinity chromatography, I cut the tag...

31 December 2014 4,945 21 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