How many neutrophils do people generally get from a neutrophil isolation protocol (using Histopaque/Dextran)?

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 View

Is it possible to remove the cells from a tissue and keep only the extracellular matrix for proteomics?

Hello! I would like to know if there is a way to flush/remove all the cells out of a tissue and keep only the extracellular matrix, in order to analyse its composition using e.g. proteomics....

14 February 2021 1,602 4 View

Meniscus formation matrigel?

Hey guys, I am currently doing tube formation assay on matrigel coated coverslips, and i wonder if there is any possibility to get rid of the meniscus formation? I have only a small part of the...

09 February 2021 6,979 3 View

Scanning Electron Microscopy for Extracellular Vesicles?

Can anyone provide or recommend a detailed protocol for preparing extracellular vesicles for SEM? Most protocols that I have found that list the necessary reagents and individual steps are for...

02 February 2021 4,475 3 View

How can I improve RNA extration from human neutrophils?

I tried several times extrating RNA from human neutrophils and the amount of RNA at the end of the precedure was minimal. I also tried using colunns from QIAGEN. Never had any problem with tissue...

26 January 2021 8,881 4 View

Why HEK293 transfected with LAMP-1GFP do not load dextran?

Hi every one I have made a cell line of HEK293 transfected LAPM-1GFP and after seeding the cells for 24h, I try to load dextran on the dish but after imaging, the cells with GFP+ have loaded less...

14 December 2020 1,801 2 View

Does anyone know the neutrophil marker for qPCR in human gut (duodenum, ileum, colon) biopsies?

Hello everyone. I have collected intestinal biopsies from stem cell transplanted patients in RNAlater and wish to quantify neutrophils only via qPCR. As we know that the mucosa of gut is filled...

25 November 2020 2,680 3 View

Which ABs concentration for EVs immuno-staining?

Hi all, I have to do a phenotyping study on cell culture derived EVs, in particular I should stain them using primary labelled ABs anti-EVs markers before proceeding to analyze them through...

25 November 2020 4,601 1 View

Can someone please share how to add a spherical wrap / correction to a visual stimulus presented to mice using python?

Hi all I use psychopy 3 to show mice a visual stimulus on an LCD screen while preforming acute extracellular recordings in vivo. I understand that one should use a spherical wrap to the stimulus...

22 November 2020 7,905 2 View

Recommend equipment of in vivo recording in mice brain?

Hello, I am trying to set up an in vivo recording system in mice brain. Which company do you recommend? Neuralynx? TDT? NeuroNexus? Which one do you guys use and why? Please let me know. Thank you!

12 November 2020 2,508 1 View

Robert C. Allen

In the past we've had consistently good yields using isopyknic focusing on a continuous gradient of colloidal poly(vinylpyrrolidone)-coated silica (Percoll).

Described in: Methods in Enzymology vol. 133 pages 459-460 (1986)

(1) (PDF) Phagocytic leukocyte oxygenation activities and chemiluminescence: a kinetic approach to analysis (researchgate.net)