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Questions related to Cell Biology
I am trying to carry out trypan blue assay. However, when I did the experiment, I was so confused about what is dead cell. My protocol : I cultured MDA MB 231 cell lines in plastic dishes. After...
07 July 2017 5,459 6 View
Hello everyone, I need to tag late endosomes, and I have FITC-Dextran of molecular weight 70KDa, can it be used to pulse chase phagolysosomes? Also how long should I treat THP1 cells with...
07 July 2017 7,047 0 View
Hi, I need to check the amount of a certain protein secreted in conditioned media. What I have done so far is plated equal number of cells and then collected equal volume of media. Following this...
07 July 2017 984 4 View
I am isolating plasma membrane from a neuronal cell line. Currently my protocol steps are as follows: 1- scrape and spin cells down at 450g for 10min. 2- Sonicate pellet in buffer and centrifuge...
07 July 2017 6,139 5 View
I would like to perform annexin assay on cultured neurons and I am afraid that accutase incubation, followed by weak centrifugation and resuspension might disrupt neurite membrane. What is the...
07 July 2017 1,627 2 View
Hi all, I was wondering if there are any limitations of growing cells or proliferation in collagen gels placed in 96 well plates. Like cell concentration, etc... Thanks in advance,
07 July 2017 2,978 4 View
Hi, I am planing to perform FISH in activate sludge samples and I am wondering if someone has worked with FISH from frozen samples prior to the fixation step. I mean, sample ---> storage (-20)...
07 July 2017 7,457 3 View
Hello everyone, I am running luciferase assay with pMIR REPORT plasmid to evaluate the function of a miRNA on a specific 3'UTR. I wondering if I could normalize the firefly signal to the number of...
07 July 2017 2,263 3 View
Hi all! I'm looking to decrease bacterial uptake by PMNs and I've been using Cytochalasin D with limited success. The bacteria are opsonized, so as far as I know this is driving a CR3-mediated...
07 July 2017 2,510 1 View
In need of a reliable housekeeping gene to normalize my pcr data to. Targeting Gfap and GLT-1 genes.
07 July 2017 8,837 5 View
co-expression about protein
07 July 2017 4,746 2 View
Which Phase of cell cycle is most radiation sensitive.
07 July 2017 2,598 1 View
I need the single M.tub to infection cells, but i find the M.tub is very easy to clumps when culture.
07 July 2017 8,774 2 View
I have recently started to study calcium and apoptosis. I have read in many reviews that "increased calcium" can trigger apoptosis, but I have not found a more detailed explanation such as the...
07 July 2017 9,728 5 View
We seem to be having trouble getting 30-70 kDa poly-L-lysine (Sigma) to remain well bound to a glass coverslip. Cells (protists in this case) won't stick if the coverslip is rinsed multiple times...
07 July 2017 9,273 12 View
I am Trying to extract high-quality RNA from acinar pancreatic cells after collagenase P digestion and FACS sorting. Can you give me some advises to obtain no-degraded RNA suitable for RNA...
07 July 2017 3,815 6 View
After removing marine cell culture media (3x L-15/ saline solution [1:1], hemolymph, and antibiotic). The purpose of the methylene blue and glutaraldehyde is to fix and stain cells, but wild...
07 July 2017 3,215 0 View
My protocol didn't work when using SD-trp culture. And nocodazole may not work. I detect the phase of yeast cells by flow cytometry. I'm looking for a new protocol for synchronization of the...
07 July 2017 4,204 4 View
We synchronize cells for different experiments, but my question is in reference to drug treatment. If we synchronize cells by minimal or no media, should we add the treatment or drug in complete...
07 July 2017 1,697 5 View
DNAase II, inhibitor, viability
07 July 2017 2,868 2 View
I am planning for an experiment using MitoTracker. I wish to do time lapse study in vitro. I want to know about the stability of the mitotracker dyes. For how long is the fluorescent intensity...
07 July 2017 8,896 3 View
I'm using HepG2 to form 3D HepG2 aggregates with alginate, to confocal microscopy analyzes. However, I have difficulty to maintaining the spheroids after fixation and all procedures of incubations...
07 July 2017 4,711 1 View
Does anyone have a protocol recommendation for digesting a collagen matrix with Cathepsin K, L, or S?
07 July 2017 2,238 2 View
Hi All, how can I induce human dermal fibroblast to necroptosis? I read some papers and it is said that different methods can induce different cells to necroptosis. Thanks.
07 July 2017 8,892 2 View