How to analyze flow cytometry PI-staining data?

More Pratik Ramjivan Shriwas's questions See All

Immunohistochemistry - How to do it?

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...

05 June 2019 9,095 4 View

Is 65% sequence identity good enough for basic modelling using Modeller?

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...

02 March 2019 7,756 11 View

Can docking results from autodock using vina be published?

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...

01 February 2019 1,276 10 View

Software for homology modelling?

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...

01 February 2019 1,762 4 View

How to removes excess blocking?

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...

31 December 2018 3,296 7 View

Crispr ko cell line?

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...

31 December 2018 9,505 10 View

How can I identify metabolites from LC-MS/MS data?

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...

11 December 2018 2,467 17 View

What is wrong with my transfer during western blot?

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...

11 December 2018 2,545 7 View

Is my blot publishable?

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...

08 September 2018 8,787 1 View

Tissue origin of HOP92 cells?

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...

07 August 2018 1,909 1 View

What is the characteristic length to determine the Reynolds number for the scattered turbulent fluid flow?

Dear all, I just saw this example in FB on the difference between laminar flow and turbulent flow. The flow from a pipe hits the sink and scattered. The flow from the pipe is characterized as...

02 March 2021 1,999 5 View

Why almost every element would be metallized in sufficiently high pressure and low temperature?

(This statement is from wikipedia, BTW). What is so special about metallic bond that is limiting high-P low-T state of a matter (as long as individual atoms exist, not white dwarf of neutron star...

02 March 2021 3,309 2 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

Why do the air flow results vary totally when adding scalar in CFD ?

Dear All, We are using Autodesk cfd to calculate the air flow in a single sided naturally ventilated room, where air flow depends only on buoyancy due to temperature difference between inside and...

25 February 2021 9,602 4 View

Unknown debris sheets observed in cell culture flasks?

Hi, We culture neural progenitor cells in T175 culture flasks coated with PLO-Laminin. We have successfully grown these cells with our protocol for months. Recently, we observed large sheets...

25 February 2021 3,443 4 View

Can I use TCA instead of TFA for protein analyis in RP-HPLC?

I am trying to analyze lipase using the reverse phase-HPLC C18 column. In the literature, 0.1 % TFA was and 60% ACN was used. Can I use TCA in place of TFA? If any other methods are available to...

25 February 2021 2,388 3 View

How to handle studies which report perfect effect sizes in a meta-analysis using Pearson's r?

I'm in the process of updating a meta-analysis and in one of the new studies, effect sizes are reported as Tau-U. Two of the effect sizes are reported as Tau-U = 1.00. I am using Andy Field's...

24 February 2021 2,265 3 View

I have need of XRD graph analysis software for phase identification. Can you please provide links for software tools which can be downloaded?

i am trying to download x'pert high score software, but can't download it. please do suggest software for phase analysis

24 February 2021 5,687 4 View

What is the error of the Txy diagram?

Hi, I know the attached Txy diagram has an error but can't figure it out. Could you please look at the diagrams and explain it to me?

23 February 2021 4,239 1 View

Why anti-CD20 or other antibodies are not used to label target cells in flow cytometry t-cell cytotoxicity assay?

New to the field of cell therapy. Why fluorescent-labeled antibodies to target cell antigens are not used for determining target cell count by flow cytometry in cytotoxicity assay (eg car-t)? Cell...

21 February 2021 3,278 3 View

Sharafaldin Al-Musawi

you can select the item in left and down of the histogram as i sign it in attached sent picture to you

Shen-An Hwang

My suggestion is for you to find someone at your university (or close by) who is familiar with the assay and know how to analyze it. Flow data analysis is not easily explained in text and off site. Especially if you are new to flow, you need someone to show you how it is done.

Diego Raffo

If the analysis is not the problem, you should check out if the staining was correct. I haven´t done it on a while, but I do remember that slowly adding the cell suspension dropwise to ethanol (while vortexing the tube) was critical for avoiding cell clumping. If you do not obtain a single cell suspension after fixation you won´t be able to see a clear graph after analysis.