Can PI (propidium staining) staining reveal the presence of mycoplasma also?

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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 5,934 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 7,392 1 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 5,056 10 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 6,263 2 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 2,606 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 1,554 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 2,508 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 9,583 3 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 9,469 1 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 461 4 View

Do I need specific antibodies to stain for a fusion protein?

I am staining some brain sections stored in cryoprotectant that express a Histone H2B- GFP fusion protein that were generated ~10 years ago. I know I need to enhance signal with an anti-GFP...

07 August 2024 5,338 2 View

How to determine positive-stained cells in FACS? Use isotype or unstained control?

To compare positive and negative cell populations in flow cytometry, should I compare unstained cells with antibody stained cells? Or with the isotype control? Most papers show comparison with...

06 August 2024 6,728 6 View

Can I multiplex a mouse monoclonal and rat primary for IHC (mouse brain tissue)?

Hi all, I was just wondering if anyone has experience with multiplexing a mouse monoclonal primary and a rat primary. I'm trying to multiplex by incubating them in the same well but was told by a...

06 August 2024 9,710 1 View

Weak DAPI staining after immunohistochemistry - how to improve?

After immunohistochemistry of previously fixed in PFA and EtOH and then frozen 20 μm sections of zebrafish brain, DAPI staining is very weak (right) compared to the same sections stained without...

05 August 2024 9,637 2 View

Senescence-associated beta galactosidase staining is False Positive in control group?

Hi guys If anyone is currently working on aging cells, you guys would like to give me some advice. I'm testing against biomarker (SA-beta-Gal), I encountered a false positive in the control group...

02 August 2024 6,735 1 View

How to correctly use the UTE and ZTE pulse sequences in Bruker's ParaVision software?

I am using a Bruker 600M solid-state NMR spectrometer with a Micro 2.5 microimaging system. The test sample is a tube of 1M LiCl aqueous solution, and the nucleus detected is 1H. I am trying to...

01 August 2024 9,227 1 View

IHC profiler in image j software. has anyone used it to quantify nuclear DAB positivity?

Dear researchers. I tried using the IHC PROFILER in image j to quantify nuclear DAB staining. I followed the instructions in the original article by "Varghese F, Bukhari AB, Malhotra R, De A...

29 July 2024 2,229 0 View

Suggestions for the Annexin V Positive Control ?

Hello everyone, I am currently using washed human platelets to stain Annexin V as a procoagulant marker. Additionally, I am staining with PerCP-CD61 to identify platelet cells. So far, I have...

29 July 2024 8,624 1 View

How can I get clearer background for my western blot?

I am running a western on HIV envelope protein. The size is around 50Kda and I am using rat anti-HA for the first stain and a second stain of mouse anti-CA. Primary antibody dilution is 1: 1000...

29 July 2024 6,484 5 View

How many events should be counted for the analysis of apoptosis with nuclear staining with Hoechst 33342 whit microscopy ?

Greeting How many events should be counted for the analysis of apoptosis with nuclear staining with Hoechst 33342 whit microscopy ? Are 100 events count sufficient and three repetitions for...

25 July 2024 7,520 0 View