23 Questions 44 Answers 0 Followers
Questions related from Mathangi Palanivel
Hi all, I am optimising ThT assay protocol for a-syn aggregation. Even if I perform 5 replicates, the graphs are having different lag times for the same sample. I am not too sure if there is a...
17 August 2023 1,883 1 View
Hi researchers, I have been running a western blot setup optimisation with beta actin as a loading control protein. I am getting non-specific bands at the higher MW range in addition to my target...
12 August 2023 6,303 5 View
Hi researchers, I am currently working on some confocal cell uptake experiments of my graphene quantum dots (GQDs), which are naturally fluorescent. I am treating SHSy5Y cells with 100ug/ml of...
10 July 2023 9,070 3 View
Hi fellow researchers! I would like to know your thoughts on the detailed steps to use cells transfected with GFP-tagged plasmid for subsequent drug studies, etc. I currently have transfected...
22 June 2023 7,330 0 View
Hi all, I am aware that EDC NHS reaction is carried out mainly for amine coupling with activated carboxyl groups. However, I have also read that it can be used for OH group coupling to form...
01 April 2023 5,038 1 View
Hi all, I am trying to differentiate SHSY5Y human neuroblastoma cells into neurons with the retinoic acid protocol. I add 10uM retinoic acid into 3% FBS containing DMEM for three days, and...
22 February 2023 8,434 4 View
Hi all, I am using the Qiagen miniprep spin kit to extract plasmid DNA. The recommended volume to culture the E. coli is 1-5mL. I have a few questions regarding my bacterial growth conditions...
09 February 2023 8,753 6 View
HI all, I have been for about a year trying to visualise my nitrogen-doped graphene quantum dots (sized less than 10nm) via TEM. Out of the many tries I have done it, I was only successfully able...
09 February 2023 1,055 6 View
Hi all, I am about to perform Lipofecatmine 2000 kit-based transfection in SHSY5Y (mammalian) cells. I have some questions regarding the minute details in the protocol, which I hope you can help...
04 February 2023 130 6 View
Hi all, I am planning on performing a Geneticin kill curve procedure on HEK293 and SHSY5Y cells before I carry out my plasmid transfection with these cell lines. Thus, I would like to put forth...
07 January 2023 5,666 2 View
Hi all, I would like to know if any of you have extracted and purified plasmid DNA without using any kit (Qiagen, Quintech, etc.), AND managed to successful transfect into mammalian cells? I...
11 November 2022 9,318 4 View
Hi, I am going to do lipofectamine transfection of my plasmid with GOI into HEK 293 and ShSY5Y cell lines. However, I have a question at the colony picking stage. How can this be done,...
11 November 2022 8,082 2 View
I am new to plasmid DNA transformation and the steps that entail, so I would like to clarify i the steps I am about to take are correct: 1. I have received my gene of interest cloned in a...
09 November 2022 9,466 6 View
I recently got plasmid from Addgene, however, I failed to expand them into glycerol stocks within 2 weeks, as recommended. It had been two months since I got the plasmid, and in the middle our 4...
09 November 2022 6,176 5 View
Hi researchers, I have 2 questions about growing SHSY5Y cells: 1. I am currently growing SHSY5Y cells in culture and just wanted to check with those who work on this cell line. Is the morphology...
25 October 2022 609 6 View
Hi researchers, I have been doing cell culture for a few months now and for the first time, I see some contamination in the MCF-7 culture that we were maintaining. I would like to know what kind...
16 April 2021 8,165 7 View
Hi researchers! We recently have bought amyloid beta 1-42 peptide from Genscript. According to the material data sheet, the peptide is soluble in water. However, after examining the amino acids,...
19 March 2021 2,489 9 View
Hi all! I have been thawing a few cell line stocks we had kept in liquid nitrogen and faced a couple of problems not being able to get the cells adhere to the T25 flask easily. I am not sure if...
28 December 2020 5,738 2 View
Hi all, I am trying to wrap my head around splitting cells into different surface-area flasks. I could really use your help! I currently have 8 T-25 flasks of HEK-293T cells. I am familiar with...
05 November 2020 3,579 4 View
Hi all, I hope you could help me out with this question in GROMACS. I am trying to use the "xpm2ps" command in GROMACS to output a postscript file from an .xpm file. However, I keep on getting...
16 April 2020 6,234 2 View
We have tried imaging the NMJs of adult drosophila legs before, whereby on anti-HRP was used as the only primary antibody. Several papers that I have read through have also mentioned anti-Dlg and...
09 May 2019 6,419 1 View
I am doing a project on trying to stain and image the neuromuscular junctions of drosophila leg muscles in great detail (with the possibility of imaging the boutons as well). So I have got two...
06 May 2019 5,718 4 View
I recently did IHC staining of mouse brain tissues with four different primary antibodies, but am unsure as to how I am supposed to assess and troubleshoot these confocal images. The following...
29 September 2017 9,593 3 View