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?
Concerned that there will be functional differences between products from Peprotech vs. Mitenyi vs. R&D.
05 June 2018 7,259 0 View
And does scraping activate cells?/ damage surface proteins on cells?
04 May 2018 4,225 0 View
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
. .
04 May 2018 6,385 0 View
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
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
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
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
For immunostaining of cells that are fixed.
06 July 2017 7,390 3 View
spun in v bottom multiwell plates
05 June 2017 2,538 0 View
Hello! I want to quantify by ELISA the secreted (from platelets poor plasma) and the non-secreted (from platelet lysate) PF4 before and after TRAP stimulation. I will use the ELISA from R&D...
03 March 2021 1,499 2 View
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
Hello! I'll do a size exclusion chromatography, but I only have an open column, and I'll perform the peptide extraction from yeast, using buffer lysis (sodium phosphate 50 mM/NaCl 30 mM/DNAse and...
01 March 2021 2,215 2 View
Can someone please give me some possible things that could go wrong? Here is my recipe: 0.5g Agarose 50 mL of TAE 1x 1 uL ethyl bromide. Gel was run at 100V for 1 hour. The buffer used is also TAE.
01 March 2021 9,952 3 View
I'm looking at the aggregation of my protein sample using DLS. Unfortunately, my buffer (20mM HEPES) also results in a set of peaks. These are at approximately 1 and 1000 d.nm. The lowest peak...
01 March 2021 9,015 2 View
Interested to stain mice brain tissue with WGA 488 conjugate, appreciate if anyone willing to offer me some guidance or suggestions! Thank you!
28 February 2021 3,951 2 View
Im doing PBMC isolation -> CD14+ enrichment using magnetic beads -> stimulation setup. My negative control is just cells in cRPMI but they seem to get activated over and over again.
28 February 2021 7,883 3 View
Hi everybody, I have been running a multiplex IHC which allow me to stain the same tissue with different markers (Primary Ab + Secondary conjugated Ab) (AF487, AF555, AF647). Since I would like to...
27 February 2021 5,028 2 View
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
Is Hoechst toxic to cells? In general, it is known as non-toxic. I am working with dsRED MDA-MB-231 cell line. I applied it for my PDMS-based microfluidic platform, for this reason, I incubated...
26 February 2021 9,119 3 View