Culture Cells

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Questions related to Culture Cells

How to check the redox state of a protein in the conditioned culture media

Hi,I was wondering if anyone knows an assay or an easy way in order to check the redox state of a protein that is secreted by the cells in the culture media?

06 June 2017 1,639 1 View

Comments on the simplest/most convenient method to isolate autologous HUVECs from humans?Comments on how easy/dificult it is to culture them in vitro?

Wondering how long these cells can survive in culture, and if cells have special requirements.

06 June 2017 5,923 1 View

Any experience with LP9 mesothelial cells' culture?

we encounter difficulties in culturing these cells. we are looking for advices how to improve cell growth

06 June 2017 3,364 0 View

Production of secondary compounds in plant tissue culture.

Hello! I would love to know about the production of secondary compounds, especially about phenolic compounds which are produced in the process of culturing plant tissue. It is supposed to be...

06 June 2017 2,265 11 View

Cellular viability

Cells HeLa grow in DMEM culture medium, 10% serum fetal bovine, 5% CO2 y 37ºC but grow is very very slow, i see a lot of black dots in and out of the cells, the dots dont are bacteria, i took a...

06 June 2017 4,362 2 View

How to check the redox state of a protein in the conditioned culture media

Hi,I was wondering if anyone knows an assay or an easy way in order to check the redox state of a protein that is secreted by the cells in the culture media?

06 June 2017 6,005 1 View

Comments on the simplest/most convenient method to isolate autologous HUVECs from humans?Comments on how easy/dificult it is to culture them in vitro?

Wondering how long these cells can survive in culture, and if cells have special requirements.

05 June 2017 3,882 1 View

After several failed attempts to measure ROS with DCFDA I am considering using mitosox. Any recommendation?

I tried to measure palmitate-induced ROS with DFCDA in H9C2 (10uM) but even though I tried several protocols (with PBS, washing prior labelling, washing after labelling, using 2X DCFDA directly in...

26 May 2017 6,599 2 View

Issues about Subcellular Protein Fractionation Kit for Cultured Cells by ThermoScientific - cytoskeletal fraction

Has anyone used the Subcellular Protein Fractionation Kit for Cultured Cells by ThermoScientific? I am very confused about the last fraction in this kit, which indicated as "cytoskeletal". But...

25 May 2017 6,087 1 View

After several failed attempts to measure ROS with DCFDA I am considering using mitosox. Any recommendation?

I tried to measure palmitate-induced ROS with DFCDA in H9C2 (10uM) but even though I tried several protocols (with PBS, washing prior labelling, washing after labelling, using 2X DCFDA directly in...

05 May 2017 2,016 2 View

Issues about Subcellular Protein Fractionation Kit for Cultured Cells by ThermoScientific - cytoskeletal fraction

Has anyone used the Subcellular Protein Fractionation Kit for Cultured Cells by ThermoScientific? I am very confused about the last fraction in this kit, which indicated as "cytoskeletal". But...

05 May 2017 6,718 1 View

Anyone has a protocol for chondrogenic differentiation of MSCs in monolayer ? and a method of quantification of the results?

I am trying to avoid the pellet protocol

05 May 2017 7,712 0 View

What could be the reason for negative OCR values in Seahorse Oxygraph?

I have started working with the Seahorse XF96 from Agilent starting with the recommended New Cell Line Characterization protocol to determine cell density and FCCP concentration. I have some...

19 April 2017 689 3 View

What is the estimated number of cells at confluence of different culture plates and culture flasks?

I wonder if anyone has an idea of the number of cells at confluency of culture plates (96, 48, 24, 12, 6-wells) and flasks (T25, T75, T175). I'm working with cartilage progenitor cells (MSC-like...

04 April 2017 1,477 3 View

What is the best method for sustaining tissue culture cells without using glucose?

I am using the pBAD24 arabinose inducible system in Shigella to express a protein of interest. My goal is to maintain induction of this protein during intracellular plaque assays. These assays...

04 April 2017 9,351 2 View

For growing Hek293 cells in MEM media, should the media have L-gluatmine or not?

04 April 2017 9,477 1 View

Can anyone suggest me an easy protocol to analysis human mesenchymal stem cell from placental tissues?

Hi , There are a lot of protocols but most of them requires culture these cells as a part of the technique , for my project i need easy protocol to preparation and analysis hMSC of placenta...

04 April 2017 4,925 4 View

Does anyone have a protocol for doing a ROS assay with SH-SY5Y cells?

I have been trying to do this ROS assay with SH-SY5Y cells but the readout is incredibly poor with little to no variation even when spiked with H2O2 if anyone has done this experiment with these...

04 April 2017 1,152 8 View

Is hypoxia induced by CoCl2 reversible?

With removel of CoCl2 in cultured cells，will hypoxic cells reagin ability to degrade HIF-1alfa protein and then become normic cells? Working on hypoxia mimics Thanks for answeing to this question.

04 April 2017 1,441 3 View

Suggestions on protocol for Cell Thawing?

Hello, Recently I thawed couple of cell lines, RPMI8226 and JJN3 and they both didn't do well during my first passage. I thawed the vials in 37 deg water bath for couple of minutes. Transferred...

04 April 2017 5,257 3 View

Why my NK-92 cells are dying ?

I have NK-92 cells from ATCC. I used two different media for cell expansion. One of them is CellGro-DC media with %5 FBS, %1 penicilin, 500 IU/ mL IL-2, Other one RPMI 1640 media with %20 FBS, 2mM...

04 April 2017 10,131 5 View

What could be the reason for negative OCR values in Seahorse Oxygraph?

I have started working with the Seahorse XF96 from Agilent starting with the recommended New Cell Line Characterization protocol to determine cell density and FCCP concentration. I have some...

04 April 2017 3,345 4 View

Can we use DMEM/MEM/RPMI culture media for MDA-MB-231 instead of Leibovitz's L-15 Medium?

04 April 2017 4,674 1 View

My media is turning yellow, but is mycoplasma negative and the incubator is fine. What could it be?

I'm culturing primary mouse astrocytes with mutant p53 in DMEM/F12 + 10% FBS. After a few days in culture, the media starts to rapidly turn yellow and the cells die. The cells are mycoplasma...

04 April 2017 7,529 7 View

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