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 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

Can somebody share a good literature on enzyme and classification of enzymes?

Hi, I wanted to know how much percentage of proteases constitutes in an enzyme family

03 April 2019 2,476 3 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

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02 March 2021 3,060 3 View

Which fluorescent dye to use for measuring total ROS production?

Is DCFDA appropriate for measuring total ROS production, or can it only detect certain types of ROS? If I want to measure total ROS production, which dye should I use?

01 March 2021 9,447 3 View

Interested to stain mice tissue with WGA 488 conjugate, anybody has a protocol to recommend?

Interested to stain mice brain tissue with WGA 488 conjugate, appreciate if anyone willing to offer me some guidance or suggestions! Thank you!

28 February 2021 3,951 2 View

No title

28 February 2021 9,936 2 View

B-mercaptoethanol (BME) protocol?

Hi everybody, I have been running a multiplex IHC which allow me to stain the same tissue with different markers (Primary Ab + Secondary conjugated Ab) (AF487, AF555, AF647). Since I would like to...

27 February 2021 5,028 2 View

Why do my MCFs not form monolayer?

Hello, I am looking for some expertise on culturing MCF7s. I purchased them from ATCC and I have been following all media and culture conditions. Media is EMEM with 10% FBS and 10ug/mL of...

27 February 2021 3,904 2 View

When is it unnecessary to change organisational culture?

As a former management practitioner, I recall a generally accepted view that changing organisational culture is difficult, so you should only attempt it if the organisational change that you are...

27 February 2021 2,595 8 View

Is there anyone who experienced with cytotoxic effect of Hoescht 33342 dye?

Is Hoechst toxic to cells? In general, it is known as non-toxic. I am working with dsRED MDA-MB-231 cell line. I applied it for my PDMS-based microfluidic platform, for this reason, I incubated...

26 February 2021 9,119 3 View

Does RNAiMAX transfection reagent work in serum free medium?

I diluted siRNA and RNAiMAX in opti-MEM and added to the cells which they were in the growth medium. Is it a right way? or should I culture cells in the opti-MEM medium for a while and not in...

26 February 2021 10,041 3 View

What is the best surface to culture cells isolated from a skin biopsy?

Hello, recently I am having some difficulties with maintaining the cell culture of primary cells isolated from a skin biopsy (normal and malignant). The coating of culture plates with Matrigel (a...

26 February 2021 6,151 2 View