Neal Edward Beeman

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Questions related from Neal Edward Beeman

Does anybody know any special tricks in detecting IFNbeta released into conditioned media?

I am using the "differentiated" THP1 cell line (PMA induction) to try to study IFNbeta response to PIC_LyoVec (a transfectable dsRNA mimetic). We get a significant response by qPCR for...

02 February 2020 10,042 1 View

Do I have to block Fc receptor when antibody staining the monocyte derivedTHp1 cell line? Does anyone have a THP1 flow cytometry staining protocol?

We've certainly got the Miltenyii reagent for blocking Fc receptor. But I don't want to use it if THP-1 cells do not need this blocking. Also, any THP1 flow tips or experiences will be well...

11 November 2019 3,245 3 View

Does anybody have a suggestion for nuclear stain to be used with pHrodoRed labeled phagocytotic probes in FACS?

I was thinking of using Propridium Iodide as a live/dead nuclear stain. The trouble is that I am already tied in to using pHrodoRed labeled Abeta peptides as a phagocytotic marker. I do not...

06 June 2019 4,701 3 View

Can I set up a plate for qPCR using Power SYBR Green PCR Master Mix #4367659 and run it a day later?

Catalog number: 4367659 Power SYBR Green PCR Master Mix. This is NOT POWER UP that I am asking about. Can I set up a plate Monday the 28th in the afternoon and run it Tuesday the 28th in...

05 May 2018 4,269 6 View

Please share a currently working protocol to harvest differentiate grow macrophages from mouse bone marrow on non adherent dishes and recombinant MCSF

I have in the past collected at the end of 7 days ~50 million macrophages from 1 mouse. I am in recent weeks getting about 10million macrophages from 1 mouse. Please include precise nature of...

08 August 2017 1,913 1 View

Are there commercial protein assays and mRNA assays for M1 Vs M2 macrophage polarization that look , preferably, at a large panel of markers?

I am growing macrophages within a synthetic matrix material that we produce. I want to compare samples from macrophages that have grown within our material to samples from macrophages that have...

07 July 2016 6,630 1 View

Does anyone have a tried and true protocol for transfecting RAW cells with plasmid DNA for protein expression?

RAW 264.7 cells mouse macrophage phenotype transformed cell line. I want to transfect RAW 264.7 cells with plasmid DNA for fluorescent reporter expression There are 2 different reporters under...

06 June 2016 8,547 4 View

Does anyone have a complete protocol for generating total cell lysate for proteomic analysis from adherent tissue cultured cells?

Can any please one post me back a protocol for generating total cell lysate from adherent macrophages (RAW 264.7 mouse cell line). I need a minimum of 10ug protein and minimum concentration of...

05 May 2016 7,449 2 View

Is there a protocol to establish a stably transfected macrophage cell line, preferably in mouse?

I need to establish a cell line that will express a bicistronic reporter. I have the reporter expression plasmid in hand. I have transiently transfected this plasmid into mouse macrophage...

01 January 1970 928 1 View

Can anyone help me to find a cell free assay or assays to compare glycolysis Vs oxidative phosphorylation?

I am growing mouse bone marrow macrophages in different synthetic scaffolds intended to support directed tissue growth and wound healing. We have shown that differences in our materials constrain...

01 January 1970 9,458 1 View