21 Questions 17 Answers 0 Followers
Questions related from Sneha Garudeswaran
Hello, I am trying to isolate exosomes from serum using SEC protocol. I have observed that the fractions mostly collected during the process are very diluted making it hard for me to quantify the...
18 September 2019 9,428 3 View
Hello, everyone, Has anyone been successful in isolating good quality exosomes using an OptiPrep density gradient? I have been trying different modifications in the OptiPrep protocol so far to...
20 August 2019 9,032 0 View
Hello, I submitted few samples (2 cell lines, 2 serum exosomes (healthy), 2 patient serum exosomes) for mass spectrometry, in order to see if we could see relevant exosome proteins and associated...
12 April 2018 7,419 6 View
Hello, I have been trying to get an optimal size for exosomes on my cell lines, but every time the size range that I obtain ends up in 160-170 nm range. I would like to perform an extra sucrose...
15 March 2018 6,912 1 View
Hello, I am working on cell lines and I make a point to validate my transfer efficiency using a Ponceau red stain, before proceeding to the blocking step. I notice that all my cell lysates show...
14 December 2017 5,349 2 View
Hello, I am trying to isolate exosomes from cell lines of DLBCL. I cultured my cells in medium with regular FBS and once they had reached confluency, I transferred the cells into exo-free FBS...
22 November 2017 4,270 3 View
Hello, Can I reuse a blot that worked previously for a specific antibody and stored at -80 degrees to test for a different antibody? The blot that I am using currently is a nitrocellulose...
11 October 2017 10,026 1 View
Hello, All, I have been trying to optimize my western blot which has been giving severe background issues since many days. I have tried to increase the wash time, even then the background...
29 September 2017 8,837 5 View
Hello, I am working on exosomes of serum and plasma and tried isolating RNA from exosomes that preserves miRNAs using the miRCURY kit. I tried assessing the sample concentration using the high...
09 September 2017 7,772 1 View
Hello, I tried isolating total RNA from serum and plasma exosomes using the miRCUry kit and submitted them to Fragment analyzer in our core facility. I was informed that they do not have a miRNA...
08 September 2017 7,572 3 View
Hello, I have been trying to validate the presence of serum and plasma exosomes using WB with TSG101 as an exosome marker since a very long time, but with no success. I use the whole cell lysates...
05 August 2017 1,562 1 View
I plan on isolating total RNA from exosomes of serum and plasma and would want to use a column based kit, since Qiagen miRNEasy kit is not fetching great results. Can I use RNeasy mini kit to...
18 July 2017 3,958 3 View
Can someone suggest on what miRNAs can be used as targets to detect healthy serum and plasma samples based on Taqman miRNA assay? I have used the spike in- ath-miR-159a during RNA isolation...
31 May 2017 1,961 1 View
We have been using miRNeasy kit to isolate miRNA from serum/plasma. Can we use the same kit to isolate miRNAs from whole blood? Inputs on this will be appreciated!
12 May 2017 4,808 2 View
I am planning to order an antibody, but I am unsure on which antibody works best as an exosomal marker for DLBCL cell lines. I have read papers suggesting the use of CD63, CD81 and TSG101 commonly...
04 May 2017 9,363 3 View
I am currently working on DLBCL cell lines, and trying to isolate RNA from exosomes of cell lines. I isolated the exosomes from cell lines using the standard protocol of ultracentrifugation and...
17 April 2017 7,128 3 View
We performed the isolation of exosomes from the TMD8 cell line growing in exo-free FBS medium after it reached a confluence of 40 million cells/10ml. We used that mediium and tried isolating...
01 January 1970 7,890 1 View
Hello, I am trying to perform proteomics on my serum exosomes, but having a really hard time doing so. The first time I could not get rid of the abundant serum proteins. The next time, I passed...
01 January 1970 3,071 3 View
Hello, I am planning to perform proteomics on exosomes as part of my project. Previously, when we had performed proteomics on few cases we only observed an abundant amount of serum proteins and I...
01 January 1970 7,418 2 View
Hello, I have isolated exosomes from serum of healthy and patient cases using differential ultracentrifugation+ultrafiltration and have validated them for their presence and absence using western...
01 January 1970 7,897 2 View
Hello, Has anyone tried the ExoQuick ULTRA kit to isolate exosomes from serum or plasma and have been successful in getting rid of the abundant serum proteins when tested against Mass Spec...
01 January 1970 8,500 1 View
