20 Questions 34 Answers 0 Followers
Questions related from Pratik Ramjivan Shriwas
Hi, I have got excellent results recently from my mice study on lung cancer xenograft tumors. I am looking to perform immunohistochemistry for analyzing the target proteins. We have never...
06 June 2019 9,183 4 View
I am trying to model a protein based on its homology. The two share 65% sequence identity. Is that good enough for basic modeling or should I opt for advanced modeling using Modeller? Btw, The...
03 March 2019 7,863 11 View
I want to perform homology modelling for proteins from GLUT family. I have GLUT1 and GLUT3 (outward open conformation). Can someone suggest any software for generating homology modelled GLUT2...
02 February 2019 1,831 4 View
Hi, I have performed a docking study using my compound with a proteins obtained from pdb wesbite. I have used autodock vina for this. Can this results be published in a manuscript. It would be...
02 February 2019 1,381 10 View
Hi, I generated a crispr ko cell line. The protein was more than 90% knocked out. But, now after several months od thawing and repassaging,the protein seems to comeback. I plan to do a grloth...
01 January 2019 9,633 10 View
I have been working with western blots and got great bands for actin. After that I stripped membrane with harsh stripping buffer but then blocked for 3hours (5% non-fat milk). I was not able to...
01 January 2019 3,375 7 View
I just performed LC-MS/MS analysis. I have got a file with m/z, retention time, fragment score, etc. I want to know how can I do a pathway analysis and identify compounds? Which softwares...
12 December 2018 2,531 17 View
I tried to transfer today and found that the membrane (membrane 1) looked like this after ponceau S staining. Two days ago, I transferred two membranes together in the same cassette and found...
12 December 2018 2,670 7 View
Hi, I am trying to blot a protein called GLUT1. It is transmembane protein known for glycosylation. So I expected to observe streak rathen than a single band. This is what I got. I tried to KO...
09 September 2018 8,842 1 View
Hi, I am working on NCI60 cell line and my anticancer-compound supposedly targets cell lines form metastatic sites (Like MDA-MB-231, ACHN, SN-12C). I wanted to know if HOP92 also has its origin...
08 August 2018 2,024 1 View
I am working on A549 cells with an anticancer compound. I am planning to study the mechanism of cell death in those cells (apoptosis or necrosis). Since, I am using flow cytometry, I want to use...
06 June 2018 7,399 3 View
I am attaching images of my cells after DAPI staining. Please tell me if they are contaminated. The problem is when I keep exposure time on microscope as 600 ms, I do not see small dots but when I...
05 May 2018 5,447 3 View
Hi, I have recently performed flow cytometry cell cycle analysis with PI. I have the FCS files and also have access to flowjo but for some reason I do not see the expected two gaussian bell...
04 April 2018 4,084 3 View
Hi, I saw lots of cells debris in my cell culture. Lots of black spots which would not go even if I washed the cells. So, I performed DAPI staining and here is my result. Can someone suggest if...
03 March 2018 8,407 8 View
Hi, I am attaching flourscence staining images with DAPI of my A549 cells. A1 and B1 are different vials. Please let me know if my cells are contaminated. I have literally wasted months...
03 March 2018 7,359 3 View
I am using fluorescence microscopy to detect apoptosis and necrosis. I use cancer cells and treat it with anticancer compounds. I am using Annexin V (apoptosis) and Propidium iodide (1 mg/ml ) for...
12 December 2017 1,076 6 View
Hi, I have a plasmid with an insert. I want to validate the presence of insert. The irony is that there is only one Restriction enzyme that cuts through the insert and it has three site son the...
10 October 2017 2,029 9 View
Hi... I want to generate a Cripsr based cell line which has three gene knockouts. Can someone share thr protocol with methods for validation at each and every step. Its like a cell expresses 4...
05 May 2017 8,413 8 View
Dear all, I want to characterize an anti-cancer drug. So, I want to test how extracellular lactate concentrations change due to treatment by my anti-cancer compound. I want to buy a kit but the...
03 March 2017 527 7 View
Hi, I have unusual western blot. My beta actin antibody is binding actin in ONE part of the western blot and the other one is binding somewhere else. I am not sure what happened. Can someone suggest
10 October 2016 5,120 3 View
