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?
Hi community, I am trying to determine what is the best way to calculate the internal consistency of a newly developed self-esteem scale. I am working together with two other researchers on the...
28 February 2021 1,369 5 View
Hi I am a bit confused. They are asking me to find out the volume of DNA required in ul (a total of 30-100 ng for genomic DNA) from the DNA concentration in the nanodrop reading which was 404.8...
26 February 2021 5,029 2 View
I am working on synthesizing and then purifying peptides using the SPPS system. After purification of the peptide, the peptide powder is obtained using a freeze dryer. When measuring the powder,...
22 February 2021 10,099 5 View
Hello, I am planning to insert GOI between CaMV 35 promoter and mGFP in pCambia1302 to produce a fusion protein and see subcellular localization of the protein. But the vector says it's membrane...
15 February 2021 4,353 3 View
Hello. I am currently trying to express a recombinant gene with Avitag at the N-terminal. I'm aiming at a final product that looks like this: Avitag-(target protein) The protein I use is already...
14 February 2021 2,039 4 View
My colleague and I are having some trouble mounting 70 micrometer mouse brain slices. When we initially position the coverslip over the mounted slices, there are no bubbles but when we come back...
10 February 2021 3,580 1 View
I am looking into DNA methylation of mice as an aging marker along with other assays and recently I added brain IF to the list of assays. For this, I need to fix the brain by transcardial...
09 February 2021 9,012 3 View
Dear all: Currently I am working with polymerization reaction in a reactor. For this, I need to control temperature, pressure, agitation speed etc. For calculation of rate of polymerization, I...
07 February 2021 355 2 View
I am trying to visualize alpha and dense granules in platelets. However, it seems like the granules were not stained well. Is there any way I can improve my staining technique to obtain better...
04 February 2021 5,925 9 View
I synthesized 1T/2H hybrid phase MoS2 on a graphite foil surface by hydrothermal method, and then analyzed it by Raman spectroscopy. As a result, three new peaks correspond to the 1T phase were...
04 February 2021 6,580 2 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