I want to stably transfect HEK-293 cells with pCMV-3 vector? What is the best method for that?
In general white opaque plates are recommended for luminescence, transparent for absorbance and black for fluorescence assays respectively. My question is if we can use black bottom plates for...
08 September 2019 5,934 4 View
I have to co-culture cancer cells with normal cells to check the efficacy to drugs target to cancer cells. In this regard, I am looking for a cell surface marker which can stain only cancer cells...
04 May 2019 7,392 1 View
I have to perform fluorescence based assays in 96-well plate using HEK-293 cells. The experimental procedure involves transfection in cells and then treatment with drugs for testing drug efficacy....
04 May 2019 5,056 10 View
I recently coated the 96 well plates with 0.1 % w/v (1:10) dilution of Poly-L-Lysine for seeding the HEK-293T cells. However, I found that the attachment of the cells to the surface is not...
04 May 2019 6,263 2 View
Hi, I wanted to know how much percentage of proteases constitutes in an enzyme family
03 April 2019 2,606 3 View
Enzymes are synthesised as zymogens and then make a transition to active form when required, is this true for every enzyme or are there any exceptions to that?
02 March 2019 1,554 4 View
I have expressed EGFP-N1 plasmid (1ug) in MCF-7 cells. Upon taking the confocal images under 40x oil I have seen the expression all over the cell? What is the possible reason for that? Can anyone...
01 February 2019 2,508 6 View
Why some cell lines are easy to transfect and some hard to transfect?
31 December 2018 9,583 3 View
Have to see protein-protein interaction in HEK-293 cells, where the donor GFP tagged protein and acceptor is RFP tagged protein. Please suggest me some good tips that I should keep in mind for...
31 December 2018 9,469 1 View
I have a vector pET23b with monomeric EGFP-6X His tagged cloned in that. The plasmid is transformed in BL21(DE3) cells. Kindly suggest me some good literature for the purification of the GFP-His...
31 December 2018 461 4 View
I have carried out MFC experiments on three different volumes, 50, 500 and 1000 mL of wastewater. Results after MFC treatment shows that TDS and EC are more in larger volumes of water i.e. TDS and...
09 August 2024 9,621 0 View
Hello everyone! I observed in my culture (htert-RPE1 cells) an orange- red particle at the bottom of the dish. It is visible to the naked eye as a very very small red dot. Could it be a...
09 August 2024 2,824 3 View
I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...
08 August 2024 4,645 2 View
I am working in fungal fermentation of soybean meal and there is bacterial growth in them at times. I am trying to quantify fungal cell counts and bacterial cells; but I haven't been able to do at...
07 August 2024 7,535 4 View
Hi, I have a question about normalizing the MTT OD values for doing the statistical analysis. So, if we have 3 different plates and we call them 3 different replicates, so, first we would...
07 August 2024 8,106 4 View
Previously when I co-coluture anti-CD19(FMC63) CAR-Jurkat with Raji with E:T=5:1, Jurkat can eliminate Raji in 24h. However, when I test another CAR construct, although I can dectect totally CD69...
06 August 2024 641 2 View
Hi! So i attempted to understand a novel protein behavior towards heat application by analyzing its secondary structure change. I subjected the protein to a thermal denaturation analysis using...
06 August 2024 1,989 3 View
Hi all, I need to introduce an ARS (autonomously replicating sequence) in my plasmid but I'm not sure which position would be the best. Does anyone have any suggestion? A picture of the plasmid...
05 August 2024 1,573 4 View
I want to introduce a point mutation (change in one nucleotide) into my gene of interest (DNA binding domain) I have designed primers as recommended on the Data sheet of the kit : -Both primers...
05 August 2024 9,059 3 View
I am performing ligation of the plasmid and a target gene. The steps I have taken are: 1. Double digestion of the plasmid and target gene 2. Ligation of the plasmid with the target gene 3....
05 August 2024 2,570 3 View