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Questions related from Sneha Garudeswaran
Hello, I am trying to set up an enzymatic assay to determine the activity of cellulase. I would like to know what sample types would be suitable to perform the same? I have read certain papers...
02 February 2020 1,219 3 View
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...
09 September 2019 5,741 2 View
Hello all, I am trying to isolate exosomes from serum and would like to know if there is some means to use the whole serum as a positive control? In my previous lab, I used to use whole-cell...
09 September 2019 5,690 6 View
I am trying to standardize exosome isolation protocol and want to run a western blot to detect TSG101. Currently, our company does not have a Tris-glycine based system and have been using MOPS...
07 July 2019 1,084 2 View
Hello, I have exosomal RNAs isolated from serum and plasma specimens of healthy and patient cases whose concentration is very low. The core facility at our university requires at least 1-10 ng to...
04 April 2019 8,613 3 View
I precipitated one of my exosome samples using the ethanol-sodium acetate process since my concentration of total RNA was pretty low. Initially, when I checked my RNA content in my exosome sample...
05 May 2018 3,923 1 View
Hello, I performed the total RNA isolation on my samples using miRCURY cell and plant kit, that also helps in preserving miRNAs. The last step of elution, asks me to elute the sample in 50 ul of...
04 April 2018 3,160 8 View
Hello, I plan on using the depletion kit on my exosome pellets, since I only see a lot of albumin and other serum proteins. Has anyone used this kit and attained success in getting rid of the...
04 April 2018 7,752 0 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...
03 March 2018 9,412 1 View
Hello, I am working with patient serum of DLBCL with starting amount of (1 ml) and isolate exosomes after which isolate total RNA enriched with miRNA. I have been successful in exosome isolation...
01 January 2018 5,707 3 View
Hello, I tried isolating total RNA that preserves miRNA from 1.2 ml of patient DLBCL serum using miRCURY kit and obtained a concentration of just 9.7 ng/ul on nanodrop. I plan on using this...
01 January 2018 9,188 4 View
Hello, I know that poly A tailing and ligation are the two additional steps used in the Taqman Advanced kit in comparison to the traditional Taqman miRNA kit. Can someone explain what is the...
01 January 2018 5,418 7 View
Hello, I isolated exosomes from DHL-16 (DLBCL) cell line, using traditional ultracentrifugation process and validated its size using Nanosight. The size obtained was within the range of typical...
12 December 2017 2,665 3 View
Hello, I always prepare fresh lysis buffer(RIPA),whenever I prepare my samples for Western Blot. However,since the volume added to my samples are very less,the rest just becomes a waste. I am...
12 December 2017 339 4 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...
12 December 2017 3,976 2 View
Hello, I am planning to isolate cell-free DNA from serum and plasma of patient samples. I would like to know what is the best kit that could be used? I came across three kits - Zymoresearch,...
12 December 2017 709 8 View
Hello, All, I am working on exosomes currently,and I have been using whole cell lysates as positive control. I always treat my cell lysates with enough amountg of RIPA buffer, estimate the...
10 October 2017 9,493 14 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...
10 October 2017 7,177 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...
09 September 2017 2,214 6 View
Hello, I was asked to perform a standard curve process using qPCR after isolation of miRNAs from serum and plasma exosomes. I normally use the Fragment analyzer to estimate the concentration of...
09 September 2017 4,480 0 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 8,039 1 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...
08 August 2017 7,388 1 View
I have completed the imaging process using chemiluminescence method and would want to preserve the blot for future reference. Can I just wrap and seal the blot and save in -20 degrees? Any inputs...
07 July 2017 9,362 3 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...
05 May 2017 6,394 3 View
I tried isolating total RNA from serum and plasma using Qiagen miRNeasy kit and estimated its concentration. The serum concentration was 0.15ng/ul and plasma concentration was 2.07ng/ul. We are...
05 May 2017 8,918 1 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...
05 May 2017 3,342 1 View
I performed an isolation of total RNA(primarily miRNA) using Qiagen miRneasy kit from serum and plasma of healthy controls and went on to perform the quantification using fragment analyzer. I got...
05 May 2017 352 10 View
I am currently working with DLBCL cell lines for my project. I have isolated exosomes from DLBCL cell line using differential ultracentrifugation and ultrafiltration after which I went on to...
04 April 2017 3,633 9 View
Hello, I have exosomes isolated from serum of patient cases and would like to use them for proteomics analysis. Previously I had used RIPA lysis buffer to lyse my exosome specimens and that...
01 January 1970 828 2 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 5,995 1 View
Hello, I am working with exosomes and wanted to see if there is any way to collect the DNA during RNA isolation. I use the miRCURY kit to isolate the RNA from exosomes that also preserves miRNA....
01 January 1970 9,835 1 View
