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

Alginate and silk fibroin hydrogels toxic to cells?

Hello, I asm currently working with alginate and silk fibroin hydrogels. I prepared a blend of alginate and aqueous silk fibroin (straight from dialysis) and resorted to ionic crosslinking to...

23 February 2021 8,249 2 View

Can anyone help me how to get the similar bands in Actin?

I run western blot on HCT-116 cell lines, I got actin band like in the attached picture. The band size is not same. Can anyone help me explain this situation and how to overcome? Thank you your...

22 February 2021 6,937 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

FDC and FRC cell populations in mouse spleen?

I was trying to isolate fibroblastic reticular cells (FRC) and follicular dendritic cells from mice spleen, but kept getting poor yield. I found from literature that the frequency of FDC is around...

17 February 2021 3,506 4 View

What to do with absence of fluorescence label in Fucci2a mouse-derived fibroblast culture?

Dear all, I have harvested fibroblast cells from R26p-Fucci2 mouse embryos (mouse line as described in this paper: https://dev.biologists.org/content/140/1/237) and cultured them on plastic,...

17 February 2021 3,368 2 View

Hi, can anybody help me with the suggestions how can I amplify a stress induced gene of 1107 bp?

I am trying to amplify the TGA1 (Transcription factor 1 ) of Arabidopsis ,which is 1107 bp. I have treated the plants with Salycylic solution over night and extracted the RNA and make cDNA. I...

17 February 2021 7,802 2 View

How to perform Kaplan-meier survival analysis in SPSS?

Hi everyone, This is my first time of attempting to run survival analysis. I have a data on patients with End-stage Kidney Diseases (ESKD). I would want to run survival probability by treatment...

16 February 2021 2,647 3 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

Patients with chronic kidney disease could get COVID-19 vaccine?

04 February 2021 278 4 View