Does company brand of cytokines affect differentiation and/or (M0/M1/M2a) polarization of TH1-P cell line into macrophages?

Concerned that there will be functional differences between products from Peprotech vs. Mitenyi vs. R&D.

05 June 2018 7,259 0 View

What does it mean when transfected macrophages increase in size (forward scatter) and granularity (side scatter)?

Cells treated only the transfection agent or the transfection agent with the miRNA construct (meant for transfection), had a higher forward scatter and side scatter compared to cells that were...

04 May 2018 1,029 1 View

Any comments on whether median fluorescence intensity (MFI) or geometric mean is better to evaluate CD marker expression using flow cytometry?

. .

04 May 2018 6,385 0 View

Any suggestions on how else I can label the nuclei of a cancer cell line that has formed a 3D cellular aggregate?

I used NucBlue Live from ThermoFisher (https://www.thermofisher.com/sg/en/home/references/protocols/cell-and-tissue-analysis/protocols/nucblue-live-readyprobes-protocol.html). I found that this...

04 May 2018 3,742 1 View

Any suggestions on a non-toxic nuclear dye for labelling macrophages while the cells are live?

NucBlue LiveReadyProbes by ThermoFisher (https://www.thermofisher.com/sg/en/home/references/protocols/cell-and-tissue-analysis/protocols/nucblue-live-readyprobes-protocol.html) seemed to seriously...

04 May 2018 5,685 3 View

Any suggestions on how I can inhibit secretion of cytokines by a cancer cell line without killing the cells?

These 'secretion-inhibited' cancer cells would then be co-cultured together with myeloid cells so I am hoping the environment would not be toxic to both cell types.

03 April 2018 5,049 9 View

Is it neccessary to use filtered pipette tips when adding lysis solution to lyse cells for downstream RNA work?

Or is the use of filtered tips only crucial from the point of isolating RNA from the lysate? (i.e. after obtaining the lysate)

03 April 2018 9,332 5 View

Can anyone share how they label monocytes with CellTracker Orange(CMRA)?

I use IMDM as staining buffer (is this ok?),but lose 60% cells after staining. Also, any idea/comment on why we should avoid amine-containing or thiol-containing buffers as the staining buffer?

11 December 2017 2,315 0 View

What markers can I use to characterize macrophage subsets (M1 and M2)? And what about different monocyte subsets (e.g. patrolling, etc)?

For immunostaining of cells that are fixed.

06 July 2017 7,390 3 View

Will I kill HUVECs when I spin/pellet them down at 700G, 4degrees?

spun in v bottom multiwell plates

05 June 2017 2,538 0 View

Waters 2707 HPLC autosampler problem?

I am using a 2707 waters HPLC device. When I try to inject a sample, it says missing plate or rack. I changed the needle and calibrated its position but I still get the same problem. I even get...

02 March 2021 1,408 1 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

How to normalise data on OPM package from R Studio ?

Good afternoon, I recently used OmniLog from BIOLOG for my experimentations : I tested the metabolism of different strains on 2 types of plates. I have 16 strains of 3 different groups...

02 March 2021 3,584 1 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

Benefits of the 24 welled plate?

We have always used the 96 welled plate, the 48 welled plate but apparently there is much evidence on the 24 welled plate and above all its benefits.It states it is the same you would use on ELISA...

28 February 2021 1,149 1 View

Can bacterial suspension be pipetted directly onto petri dish and agar poured onto it?

Hi, recently I encountered a protocol for plating which was not the usual method I have been practicing (pour LB agar onto plate, allow it to solidify, pipette bacterial suspension onto plate,...

28 February 2021 7,292 3 View

What is the best surface to culture cells isolated from a skin biopsy?

Hello, recently I am having some difficulties with maintaining the cell culture of primary cells isolated from a skin biopsy (normal and malignant). The coating of culture plates with Matrigel (a...

26 February 2021 6,151 2 View

Difficulty in cloning of 1kb gene into vector?

I am facing difficulties in cloning a 1kb gene into a vector (pJIT163). I have my gene on interest (GOI) in pUC57 and want to clone in pJIT163 using SalI and BamHI restriction sites. I am getting...

25 February 2021 3,221 7 View

Can cells be reused after using WST 8/ CCK 8 from Sigma Aldrich?

Hi all, Can cells be reused after I have seed them onto 96 well plates and test them with cell counting kit at OD450 (CCK8) at 12 hours, wash them with PBS x3 and add new media and let them...

24 February 2021 5,072 1 View

Standard curve shifting over time. How do I stabilise the ELISA?

24 February 2021 1,365 3 View