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Questions related to Culture Cells
I have been trying to isolate exosomes from HEK293 cells. and i have read and tried several protocols. but still can't get any exosomes pellet in the final step. How long should i culture cells in...
10 October 2019 8,164 5 View
Hi everyone. I am working on primary culture of OSCC. After 25 days of culture cells have grown like these. Can anyone help me in identifying the type of cell please?
10 October 2019 1,967 3 View
09 October 2019 2,724 5 View
I came across some protocols using Formic Acid in the first step of Amyloid beta extraction from cultured cells. Formic Acid is not the safest substance to use, so I was wondering if there is a...
10 September 2019 6,034 1 View
I need a straight protocol to study deposited ECM in cultured cells upon stimuli with selected molecules
09 September 2019 1,030 2 View
I have successfully cultured nasal polyps epithelial culture cells using BEGM medium with streptomycin and fungizone, in T25 flasks but i have problems while growing from brushes and i also have...
09 September 2019 3,709 2 View
I have observed that when I culture cells or organoids on a petri dish that are placed in an incubator (with humidity, 37 C and 5% CO2) on an orbital shaker at 80 rpm, there is more water...
28 August 2019 563 3 View
I have an XY cell line I am measuring telomere length on, and I am using an XX cell line (with abnormally long telomeres) as an internal control. I would like to identify each separately, but that...
27 August 2019 1,460 1 View
Hello I have cultured cells and I want to extract proteins before using the PNGase for deglycosylation and SDS PAGE. The protocol from the provider for the PNGase is pretty simple, but what...
29 July 2019 9,060 2 View
I am looking for a tool or methods which can help me to choose the stretch of sequence from my target miRNAs which can be used to design siRNA in order to inhibits its expression in cultured cells.
15 July 2019 5,882 2 View
I need to detect ROS in cultured cells like Jurkat, HL-60, with H2-DCFDA reagens by fluorescence microscope and don't have a protocol. Also, I saw on the internet that phenol red should be...
07 July 2019 5,401 1 View
I'm in shock right now! I checked my flasks this morning and found strange things init! This is my first time finding like this in my culture!! Although I follow a precise aspect technique in my...
07 July 2019 8,054 1 View
I have tried isolating PBMCs (looking to culture monocytes and differentiate into macrophages) from human blood using Ficoll gradient. I get a good amount of cells that attach. After 2-3 hours I...
07 July 2019 938 6 View
The RNA concentration in serum according to literature is about 5-40 ng/ml. However, there is very little information about the RNA concentration in conditioned medium of cultured cells....
28 June 2019 6,575 7 View
I have started using the Thermo Fisher Heracell 240i tri-gas incubator for culturing the cells in hypoxia (5% Co2 and 5% O2). I have connected the incubator to nitrogen gas cylinders to achieve...
06 June 2019 7,650 2 View
Hello guys I intend to label mRNA sperm with ISH technique but couldn't find a relevant protocol. What do you think if I use ISH protocol used for cultured cells? I appreciate your feedback, in...
06 June 2019 3,510 2 View
06 June 2019 868 5 View
I am trying to prove that some of the in vitro effects I observe following exosome treatment in target cells are miRNA dependent. In this phase I do not want to focus on a specific miRNA but just...
26 May 2019 6,126 5 View
Is it best perform fixation step with formaldeihyde or paraformaldeihyde at 4ºC or room temperature or there is no great difference? I work with human mo-DCs in culture and I need to detect...
13 May 2019 3,774 3 View
Has anybody ever used azole antifungals (fluconazole etc) in their cell/tissue culture? I work with primary human bone/cartilage tissue and I struggle with amphotericin B resistant fungal...
05 May 2019 2,592 1 View
05 May 2019 7,022 6 View
Hi, there, I am doing primary culture of colorectal cancer. In recent months my cultures are contaminated. I suppose it is some kind of bacterial. It grows slow and almost always appears after 4...
23 April 2019 2,238 3 View
04 April 2019 6,808 3 View
I am culturing the cells (isolated from bone marrow) after treatment with RBC lysis buffer with the aim to have a CD11c+ DC population at the end of 7 days. I am using 10ng/ml GM-CSF in RPMI-1640...
04 April 2019 5,731 6 View