Can black bottom 96-well plates be used for both fluorescence and luminiscence assays?

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 5,751 4 View

Can anyone suggest a cell surface fluorescent markers (dye or stain) that can exclusively stain cancer cells and not normal cells ?

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 7,244 1 View

Can anyone suggest me a better coating reagent other than Poly-L-Lysine for the adhesion of HEK-293T cells?

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 6,056 2 View

How to perform cell based assay in 96-well plate using HEK-293 cells as the cells shows poor adhesion and comes off quicking ?

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 4,877 10 View

Does all enzymes are synthesised by ribosomes in zymogen form or are there expections when enzymes are synthesized in active form?

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 1,388 4 View

Why am I seeing expression all over the cell while expressing EGFP-N1 plasmid in MCF-7 cells?

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 2,337 6 View

Why HEK-293 cell line offer relatively high transfection efficiency as compared to other cell line?

Why some cell lines are easy to transfect and some hard to transfect?

31 December 2018 9,414 3 View

Which cell line offers the best transfection efficiency?

Transfection efficiency depends on a lot of factors among which the ''the cell line type'' is one of them? In general, which cell line shows maximum transfection efficiency and how much?

31 December 2018 3,338 6 View

I have to purify a GFP-His tagged protein. Kindly suggest me some good literature in this regard ?

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 309 4 View

I have to perform FRET assay using a microplate reader in a 96 well plate. Please suggest me some good literature to perform the assay?

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 9,345 1 View

How to calculate the fraction metabolized of a drug by an enzyme using clearance(ml/min) values of the metabolite formed?

I am trying to find the fraction metabolized of a drug by CYP and UGT enzymes. I am unable to get the values of fraction metabolized (Fm) directly from the literature. Can we calculate the Fm of a...

21 February 2021 5,379 2 View

How long does it take to design and validate transition state analogue inhibitors?

Hi, I'm asking from the point of view of designing a research plan: How long (approximately) can it take to determine the transition state structure of an enzyme substrate using kinetic isotope...

14 February 2021 2,610 1 View

Kinetics of Enzyme production in Recombinant E Coli?

Hello, I am trying to find kinetic information (such as rate of enzyme production etc.) on the expression of taq polymerase in E-Coli after inserting a recombinant plasmid. Any help would be...

12 February 2021 1,386 3 View

No title

Hello! I want to determine the activity of my in house Reverse Transcriptase enzyme. For this, I have used Thermo's Enzcheck Reverse Transcriptase assay kit. I used commercial Thermo RT enzyme to...

08 February 2021 7,285 1 View

How define one baker unit(BU) is the amount of hydrogen peroxide catalase ?

07 February 2021 2,006 1 View

Does DNA A-tailing survive after agarose gel electrophoresis and purification?

Hi everyone, I need to clone in pGEM-T vector the result of amplifying a metagenomic library by PCR with a high fidelity polymerase but I only want to clone the fragments of an specific size...

04 February 2021 6,051 8 View

What are some common oxidants used to oxidize iron sulfur clusters in proteins for techniques like EPR?

I have used potassium ferricyanide (which is likely the most common) extensively in the past, but I am curious to know if there are other oxidants that work well. I need to generate the oxidized...

29 January 2021 9,803 4 View

Papain Enzyme Assay...?

Hi all, I would like to know how can I estimate the activity of the enzyme, papain, which is used in reactions as crude latex extract of papaya. Please share some procedures if anybody having...

26 January 2021 569 4 View

Where can I find plants pyr-GC-MS data?

25 January 2021 5,403 2 View

Recombinant restriction enzyme: how strong should the corresponding methyltransferase expression level be?

Hello. We are planning expression of a restriction enzyme in E. coli, and wondering how much of the corresponding methyltransferase is needed to be expressed in order to protect the host genome...

23 January 2021 2,096 4 View