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

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

Can we classify HV and LV lines based on line length?

I have dataset which shows the length of power lines. I need to classify the lines based on the line length. Is there a rule to classify the High voltage (HV) and low voltage (LV) lines based on...

03 March 2021 4,116 4 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

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02 March 2021 3,060 3 View

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 View

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 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

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

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 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