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Questions related from Tomasz Maciej Stępkowski
Redox cycling agents such as menadione or paraquat can "obtain" one electron by reduction by various enzymes such as: NADPH-cytochrome P450 reductase (E.C. 1.6.2.3) , NADH-cytochrome b5...
26 June 2018 5,094 2 View
In http://www.sciencedirect.com/science/article/pii/S1383574215000022 the authors review some articles showing that chemo and radiotherapy can introduce new heteroplasmic variation in mtDNA. Is it...
06 July 2017 8,994 0 View
I used to grow my undifferentiated LUHMES cells (dopaminergic neurons precursors) in 75 cm2 bottles and then add differentiation medium for 24hrs and seed them into four 21 Cm2 bottles and grow...
29 October 2014 4,484 2 View
I have a little time for optimization and therefore will be happy if someone could provide an experimentally validated sequence of primers for RT-PCR amplification of human housekeeping gene...
05 August 2014 1,688 2 View
Dear I recently tried SURE FIND(TM) transcriptomic arrays from SA Bisciences (http://www.sabiosciences.com/transcriptome_pcr_product/HTML/TCSC-601A.html) that contain cDNA from siRNA treated cells...
30 July 2014 1,381 3 View
Can anyone tell my if my calculations and assumptions make sense. My gene of interest got two isoforms and I want to found out how the proportions between those isoforms change in various...
24 July 2014 2,950 3 View
Could anyone help me with choosing a proper concentration and time point to study gene expression changes after treating dopaminergic neurons (differentiated LUHMES cell line) with 6...
17 July 2014 3,258 2 View
What is the reason for such an irregular band shapes in Western Blot (Narrow in the center with bulks on the border). I sometime got those and I am not sure what is the reason. I routinely use...
10 April 2014 3,054 13 View
I coat my cell culture flasks with a mixture of poly-L ornithine and fibronectin overnight then aspirate the coating solution and dry it open in a laminar flow cabinet under UV light . Is the...
03 February 2014 7,155 5 View
1. Can anyone explain to me what to expect when comparing the overexpression of the gene and subsequent protein from lentiviral plasmid during transient transfection and transduction. Are the...
13 December 2013 6,267 8 View
Does anyone know a paper in which researchers created a real image or computational model of mitochondria dynamics in 3D in time. The papers discussing the problems of such analysis and image...
30 October 2013 4,225 3 View
I am curious what is the extent of the alternations in mitochondrial network structure after cell culture fixation - is it negligible or not? What is the best protocol for fixation of cells to...
29 October 2013 8,158 2 View
We are going to upgrade our NIKON A1 confocal system with the incubator for long lasting live cell imaging - the incubator should contain atmosfere humidifier, CO2, temperature control etc....
15 October 2013 1,400 7 View
I performed a Real-Time PCR expression profiling for 85 NF-kB pathway related genes. In my experiment conditions some genes show high, little or medium statistically significant changes in...
18 June 2013 4,371 15 View
Can anyone tell me if the PGK promoter driven expression of mitoDsRed2 will be strong enough to visualise mitochondria in living cells and sort mitoDsRed2 expressing cells in FACS? Do I need a...
03 June 2013 1,002 3 View
I would like to create lentiviral derived transduced cell lines (neuronal and lung cancer) with overexpression and shRNA knockdown of my protein of interest. As I am going to study mitochondria...
14 May 2013 9,482 3 View
Has anyone performed Western Blot comparison of protein abundance during neuronal differentiation? I would like to show the difference in expression of my protein during the differentiation of...
07 March 2013 8,629 18 View
Please tell me what the latest research says about what kind of diet is beneficial and healthy in the prevention and treatment support of joint degeneration especially in the case of young men...
04 March 2013 7,688 0 View
I want to design a fusion of GFP or YFP to the C- terminus of my protein. I am doing it for the first time so please help me with some golden rules - 1. What amino acid composition and length is...
01 March 2013 3,231 5 View
Can anyone suggest good companies where I can buy custom made lentiviral plasmids for my gene of interest - I would like a construct with conditional (on, off) expression with and without GFP tag....
28 February 2013 10,084 23 View
Protein lysates are more stable when you quick freeze them in liquid nitrogen. To my knowledge during slow freezing a density gradient is formed that can lead to protein precipitation and...
21 February 2013 8,169 1 View
Please help. I am planning to do a Western Blot LC3 turnover assay to study autophagy flux in an A549 lung cancer cell line. I have only got leupeptin protease inhibitor in the lab. Is it...
25 January 2013 6,045 7 View
I got an alignment of orthological promoter sequences of 13 eutherian mammals created in ENSEMBL.org. I have found some motifs that are relatively well conserved among tested mammals and proved...
22 January 2013 2,738 2 View
Does anyone know if it is possible to achieve sub cellular (nuclear, mitochondria, cytoplasm) enriched fractions from frozen cell pellets? If so please help me with the protocol and appropriate...
26 November 2012 5,540 10 View
I am going to use Phylogibbs program to study potential TF binding sites in 7 orthologous promoter regions derived from the known genomes of human, chimpanzee, rhesus monkey, rat, mouse, cattle...
28 January 2009 929 1 View
