Suitable sample types for assaying cellulase activity?

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...

01 February 2020 1,105 3 View

Positive control for exosome isolation from serum?

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...

08 September 2019 5,582 6 View

Which running buffer is more compatible to detect TSG101 for exosomes?

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...

06 July 2019 969 2 View

Exosome miRNA sequencing core facilities?

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...

03 April 2019 8,504 3 View

Does ETOH-NaOAc precipitation on RNA samples isolated from exosomes increase the ribosomal RNA content?

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...

04 May 2018 3,805 1 View

Can we concentrate the RNA that was eluted in elution buffer?

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...

03 April 2018 3,047 8 View

Did someone use the "High Select Top 14 Abundant Protein Depletion Mini Spin Columns" for depletion of abundant proteins and were successful?

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...

03 April 2018 7,632 0 View

Working protocol for sucrose cushion?

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...

02 March 2018 9,284 1 View

How to increase the concentration of RNA?

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...

31 December 2017 9,060 4 View

Why is the poly A tailing and ligation step necessary in Taqman Advaced miRNA kit?

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...

31 December 2017 5,297 7 View

Buffer Exchange using column packed with sephadex G-25 effect on protein concentration?

Hello, If i am doing a buffer exchange for an antibody of 1mg/mL, does the elution lose protein in the process of buffer exchange? For example, if i flow through 500 uL of 1mg/mL sample, and...

03 March 2021 6,299 3 View

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03 March 2021 5,360 2 View

Sample dilution for quantitative analysis using ELISA?

If the detection range is in ng/ml but the reference range is in ug/ml for a molecule or protein in serum or plasma .how to dilute and what is the initial volume to be taken for quantitative analysis

02 March 2021 7,670 3 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View

What is the most suitable server for predicting protein structures in bacteria?

I have used the i-Tasser several times, however it has been unavailable for several days. I tried the swiss-model, but the output was not very pleasant due to the model used. Is there any other...

28 February 2021 4,521 3 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

Are protein samples in SDS compatible with ELISA?

I have used an AllPrep DNA/RNA/Protein Mini Kit QIAGEN kit to extract RNA and protein from my samples. At the end of the protein purification, I resuspend my protein in 5% SDS. Will these samples...

28 February 2021 7,370 3 View

Anyone that can help with details in phosphopeptides enrichment?

Anyone know the amount of protein before enzymatic digestion for phosphopeptides enrichment of serume sample in Ti-IMAC? Has anyone ever use Ti-IMAC(the material is from Chinese company named...

28 February 2021 7,310 3 View

Does anyone use antinuclear antibody (ANA) and anti-Phospholipid antibody ELISA kit for Non Human Primate serum samples?

I am looking for anti-nuclear antibody (ANA) and anti-Phospholipid antibody ELISA kit for Non Human Primate serum samples. Any suggestions highly appreciated. Thank you.

28 February 2021 6,512 3 View