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Questions related from Puneeta Bathla
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...
09 September 2019 5,895 4 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....
05 May 2019 5,012 10 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...
05 May 2019 7,351 1 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...
05 May 2019 6,219 2 View
Hi, I wanted to know how much percentage of proteases constitutes in an enzyme family
04 April 2019 2,560 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?
03 March 2019 1,505 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...
02 February 2019 2,457 6 View
I have to do some bioluminescence assays, in that regard if anyone has a caspase-3 or any other caspase-luciferase construct, kindly let me know.
01 January 2019 4,289 3 View
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?
01 January 2019 3,450 6 View
Why some cell lines are easy to transfect and some hard to transfect?
01 January 2019 9,532 3 View
I have purified a EGFP-His tagged protein which was there in pET15b vector using Ni-NTA column. The concentration obtained was way too high around 5mg/ml. Before the running the SDS-PAGE the...
01 January 2019 4,937 3 View
I have purified a monomeric EGFP- His protein with a molecular weight of 30 kDa. But, instead of 30 kDa I am getting a thick band of 60 kDa in SDS-PAGE reducing gel. what is the possible reason...
01 January 2019 6,924 2 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...
01 January 2019 423 4 View
I have transfected a GFP tagged plasmid in HEK-293 cells in a 96-well plate. I am not able to see the difference in reading in un-transfected versus transfected cells using a microplate reader...
01 January 2019 7,627 3 View
Best buffer composition for storing a recombinantly expressed and purified mEGFP protein ?
01 January 2019 1,973 4 View
I would be doing a BRET assay for the first time. What are the key controls and points I should keep in mind for performing a BRET assay?
01 January 2019 2,464 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...
01 January 2019 9,439 1 View
I am looking for a luciferase reporter stable cell line?
11 November 2018 4,920 1 View
Hi, I need to do some preliminary BRET experiments and for that I am looking for a luciferase stable cell line. Kindly let me know if anyone has it. Thanks!
11 November 2018 2,521 1 View
As PFA fixation is known to denature proteins and GFP fluorescence is highly dependent on its structure, so would PFA fixation will hinder my imaging experiment where I have a GFP tagged protein...
10 October 2018 5,548 5 View
FLAG tag the octapeptide sequence and artificial epitope binds to the FLAG antibody. I want to know in which region of the FLAG antibody does the FLAG epitope binds ?
09 September 2018 4,206 2 View
I am looking for a commercially available fluorescent molecule which is uncharged
09 September 2018 3,373 3 View
My data set has lot of confocal imaging data.. what software should I use for processing it for a publication purpose?
07 July 2018 3,698 0 View
I want to track apoptotic events in HEK-293 cells and specially the time-period when caspase-3/7 gets activated after apoptotic induction.
07 July 2018 1,088 0 View
Major good writing tips for writing a scientific paper?
07 July 2018 7,038 11 View
I'm looking for a protocol for staining mitochondria using TMRE dye?
07 July 2018 7,524 3 View
Looking for the following cell line described in the question? Kindly let me know if anyone has it?
06 June 2018 6,556 0 View
I have a protein of molecular mass 400 kDa? What is the required protein gel concentration for both stacking and resolving gel?
04 April 2018 447 1 View
My current experiments involves drug screening, in this regard I am looking for a list of cyto-toxic drugs with unknown mechanism of action.
04 April 2018 5,092 2 View
I am trying to see the effect of a drug on cell viability. The drug was dissolved in sodium phosphate buffer (50mM). In control cells where no drug was added but the buffer was added, a lot of...
04 April 2018 8,312 1 View
I want to stably transfect HEK-293 cells with pCMV-3 vector? What is the best method for that?
03 March 2018 7,201 0 View
I would like to know if anyone has created a stable cell-line of MCF-7 cells expressing Caspase-3 GFP gene?
03 March 2018 1,391 1 View
I've got a plasmid from SinoBiological- Casp-3 GFPspark cDNA clone. In the section of expression method it says- Constitutive, Stable/transient. I would like to know what ''stable'' is here? I...
03 March 2018 8,989 2 View
I want to stably transfect MCF-7 cells with caspase-3 GFP plasmid. I've not performed stable transfections in the past. So, I'm looking for a protocol which will help me to do that.
03 March 2018 8,046 8 View
I have to perform an intracellular FRET assay in 96 well plate and looking for a nice protocol in this regard.
03 March 2018 1,789 1 View
I want to know some useful tips which help me in doing transfection in 96 well plate ?
02 February 2018 8,945 3 View
All enzymes undergoes post-translational modification (PTM) to become a functional mature enzyme. Is there any enzyme which doesn't undergo PTM? any exceptional enzyme?
02 February 2018 9,364 3 View
I am in need of a cell line which is deficient in caspase-8 gene. Please let me know if anyone has it? or any details regarding the particular cell line.
01 January 2018 5,643 1 View
I made a stock solution of 500uM for Z-VAD-FMK caspase inhibitor using DMSO solvent, however, I now realize that I need a stock of either 1or 5mM. Is there any trick to evaporate DMSO solvent in...
01 January 2018 3,010 2 View
Nuclear stain like DAPI and Hoechst stains nuclei and also reveals the presence of mycoplasma. So, my question is does PI stain along with staining the nucleus tell us the presence of mycoplasma?
01 January 2018 8,160 2 View
I have kept the eppendrof tube in laminar hood for 10min. Can UV-B light (laminar hood) penetrate inside the eppendrof tube?
01 January 2018 456 0 View
Studying necroptosis pathway
12 December 2017 612 2 View
MCF-7 cell line is caspase-3 null that means it doesn't express caspase-3 and hence a suitable model for studying the function of caspase-3 by exogenous expression tagging it with fluorescent...
12 December 2017 6,437 4 View
I'm trying to transfect MCF-7 with 2ug caspase-3 GFP plasmid using Lipofectamine2000 and the transfection efficiency is terribly low. Can anyone suggest me something which can enhance the...
12 December 2017 2,546 8 View
Lipofectamine Stem reagent (Invitrogen) on the website it is mentioned that the reagent is for transfecting stem cells with low toxicity. So can I use this for transfecting MCF-7 cell line?
12 December 2017 1,894 2 View
