Mixture neutralization after Boc deprotection ?

I am performing a Boc-deprotection with HCl in organic solvent (dioxane or Ethylacetate) but would like to isolate the free amine. Several problems: - Neutralizing with inorganic base as NEt3...

18 February 2021 3,366 3 View

Murine P2X7 western blot?

Hello, I am trying to do western blots of murine P2X7, without success so far. Does anyone know a good antibody for western blotting of P2X7 from murine cells? Thank you very much in advance.

07 December 2017 737 3 View

I have interesting data about bilateral deficit hand dominance but i don't want to work alone. Is there anybody interested by a collaboration ?

11 May 2015 3,007 3 View

What could be some reasons why unstimulated monocytes are getting activated?

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

Looking for advices and tips to improve the staining and the gating strategy in human blood?

Hi, I am staining a human blood with CD19, CD3,CD14 and CD16. My gating strategy in FlowJo goes like this: - First, I gate a general population SSC-A VS FSC-A => Then gate for single cells, =>...

21 February 2021 5,724 4 View

Will SARS-CoV-2 Spike be broken into S1 and S2 by denaturing conditions?

Hello, I am performing a western blot of my SARS-CoV-2 Spike-pseudotyped lentiviral particles. I denature my virus by adding concentrated virus to 2X Laemmli buffer at 1:1, then boiling for...

17 February 2021 4,844 4 View

Macrophages in culture not growing/ dying or detaching . Why ?

Hello, I am an undergraduate student doing my dissertation, where I isolate PBMCs from leucocyte cones in order to plate monocytes and allow for macrophage differentiation. Although the numbers...

09 February 2021 5,334 8 View

Will it be possible to get IC:50 if we target cancer cell line with antibody drug conjugate (ADC) that have 50% less antigen positive cells ?

Is it possible to attain IC50 if my target cells have less than 50% antigen expression ? For example: Cancer cell line XXX is showing 35% of expression of cell surface antigen YYYY, if i...

04 February 2021 2,711 3 View

What is the best concentration of Pam3CSK4 to coat a dish to activate with THP-1?

Hi everyone I am trying to coat a glass-bottom dish with Pam3CSK4 and plate THP-1 cells to measure IL6 with ELISA from the supernatant. I wondered if anyone knows a good concentration and...

21 January 2021 2,593 1 View

Is there a reason why my monocyte population had high CD3 expression?

We ran a flow cytometry experiment where Fc blockers were added into the antibody cocktail. Other antibodies added include: 1. anti-CD3-PE-Cy7 2. anti-CD4-APC 3. anti-CD8-AmCyan 4....

10 January 2021 2,806 4 View

Does CD31 indicate cell viability?

Hi, I am wondering if the immunofluorescent staining of CD31 antibodies can both mark endothelial cells and indicating their viability. In other words, if I perform immunofluorescent staining for...

23 December 2020 5,815 4 View

PDX leukemia in immunodeficient mice show regression after a time. Anyone knows what is going on?

I am performing PDX ALL in NSGS mice, but something curious happens: some leukemias develop very well , but after a time the hCD45+ cells show a huge regression while the mCD45+ cells turn back to...

16 December 2020 7,344 2 View

IHC-P staining of the viral infection in the mouse pancreas?

Hi, I'm trying to stain a viral infection of the pancreas in paraffin sections of adult (3 month old) BALB/c mice. Prior to starting my histology analysis, I have titrated the organ to confirm...

10 December 2020 2,198 3 View