How best to thaw plasma samples for an immunoassay (cytokines)?

When I defrost my plasma samples from -80, I do so on ice, slowly. Then I centrifuge the samples. Upon centrifugation, the plasma samples will have a white sticky substance layer at the top, but...

30 July 2020 4,877 2 View

Different RNAdatase comparison?

Hi, I would like to compare my dataset generated with Illumina system with other datasets published and deposited on GEO. Is free software that allows me to do that? or just R software is enough?

29 September 2019 3,356 3 View

How can I clean glassware from TiO2 nanoparticles?

In my lab we've been synthesizing 10-20 nm TiO2 nanoparticles from Titanium Isopropoxide, and now the glassware, no matter how I clean it with solvents, is marked with a white patina. I could have...

05 May 2019 5,747 10 View

Influenza viruses plaque assay ?

Hi, I want a correct protocol for plaque assay with influenza viruses . I used MDCK CELLS ( 450 000 cell/well ) with different strains and different plates but i didn't see plaque formation but...

17 February 2019 5,686 4 View

Can protein folding affect antibody detection in a western blot?

I have problems with my western blots in that I get no signal. I have tried all the usual changing of ab concentration, different blocking etc but nothing is working. Is it possible that my...

21 October 2018 5,207 6 View

Have anybody here performed an abductive research strategy as a tool for textual interpretation of interviews?

Do you have any concrete suggestes about the methodological tools to handle abduction? I find that sorting an categorizing the data by coding is not satisfactory, and are looking for an...

01 August 2018 3,983 7 View

Is it possible to use regular Tris-Glycine Running buffer (SDS) on a NuPAGE Tris-Acetate 3-8% precast gel?

15 August 2017 8,144 3 View

THP 1 migration

Hi everyone! Do you have any suggestion regarding THP 1 migration assay? How many cells do you plate in a 24 well transwell with 8 um pore size? Incubation time? Thanks!

08 June 2017 6,333 2 View

Protocol for F4/80 IHC on lung samples?

Hi everyone! I am trying to perform IHC for F4/80 (ab6640 abcam) on pfa fixed LUNG samples, but i'm not able to obtain a good staining. Do you know a good protocol? I usually use a concentration...

20 February 2017 2,652 2 View

What is the best method to concentrate PCR product after pooling several samples?

28 June 2015 5,116 11 View

Should we change media after1st 6hrs after plating HUVEC bought from INVITROGEN?it's not recomended by invitrogen.Will DMSO present in stock matter?

We have HUVEC bought from Invitrogen. Cat no -C-003-5c. while first plating the cells we have bought the protocol suggests not to centrifuge the cell with media to remove the DMSO. Should we...

02 March 2021 3,713 2 View

Bought HUVEC from Invitrgen.need to revive it.expected it is cryofreezed in DMSO.Do I need to remove the DMSO for plating the cell for the 1st time?

After I cryofreeze the cell do I need to remove the DMSO for culturing the cell for next passages .

28 February 2021 7,738 3 View

For gelatin coating for culturing HUVEC.Should the gelatin be made in sterile water or PBS? Do I need to autoclave the gelatin soln after making it ?

I want to use 0.1% gelatin. Should that be made in PBS or sterile water? Should I autoclave the gelatin soln after making it in water/PBS?

28 February 2021 274 2 View

Why do my MCFs not form monolayer?

Hello, I am looking for some expertise on culturing MCF7s. I purchased them from ATCC and I have been following all media and culture conditions. Media is EMEM with 10% FBS and 10ug/mL of...

27 February 2021 3,904 2 View

Help with culturing monolayer of human brain microvascular endothelial cells (hBMEC)?

I've been having a hard time forming an endothelial monolayer with good tight junctions. The person who trained me on the culture of endothelial cells worked mostly with mouse BMEC, and he used...

22 February 2021 8,253 1 View

Meniscus formation matrigel?

Hey guys, I am currently doing tube formation assay on matrigel coated coverslips, and i wonder if there is any possibility to get rid of the meniscus formation? I have only a small part of the...

09 February 2021 6,979 3 View

C-4 I cervical cancer cell line tissue culture protocol?

Hi there! My lab has been brought an array of C-4 I cell line. The cells did not succeed in attaching to the bottom of the flask and remained floating in the media. Although we followed the...

04 February 2021 10,018 5 View

Cell Culture Protocol for 67NR and 4T07 cells?

Hello there! My lab has recently been (very graciously) gifted an array of cell lines, of which include 67NR and 4T07 cell lines. While I've found a decent amount of background and culturing...

02 February 2021 2,590 3 View

Moving bacteria-like organisms in my HEK293T cell culture?

Hi everyone, I have been culturing HEK293T cells that have been engineered to overexpress the mechanosensitive protein Piezo1. However, I am finding it very difficult to keep them in culture -...

02 February 2021 7,892 4 View

How do you quantify bacterial qPCR data?

Hi all, How would you use qPCR to quantify the amount of a particular bacterial species in a sample experimentally? I understand you could use specific primers for the 16s V region of a...

01 February 2021 7,153 4 View