For parallel storage of proteins, RNA, and DNA in their cellular context
CellCover was developed for fast “one step” stabilization of biomolecules. Its non-toxic formulation instantaneously protects and preserves RNA and protein expression patterns in human and animal cell types. This includes cells found in solid tissues, such as tumors, and cultured cell clusters, such as organoids and spheroids. Labs can then analyze the treated cell’s DNA, RNA and proteins days or weeks later with almost no detectable change in levels, patterns or modifications.
The rapid fixation of RNA and proteins is very important for expression patterns to remain unchanged. With other techniques, such rapid fixation is often at the expense of other factors such as molecule crosslinking by PFA (paraformaldehyde) or cell lysis by high-salt reagents that stabilize RNA. Not so with CellCover, which instantly fixes and stabilizes cells in their true state — maintaining RNA and protein expression.CellCover, which instantly fixes and stabilizes cells in their true state — maintaining RNA and protein expression.
As soon as cells are exposed to CellCover their metabolic state is “frozen in time” without actually freezing the cells. This chemical-induced freezing instantly stalls all cell processes including: i) synthesis and processing of bio-molecules, and ii) disruption of cellular degradation pathways. In particular, its inhibition of chemical and enzymatic degradation leads to exceptional stabilization of DNA, RNA and protein molecules. CellCover is the perfect choice for for analyses that are looking for a “snapshot” in time, e.g. flow cytometry, mRNA expression profiles, NGS, and protein post-translational modifications.
Preserves morphology, mRNA expression patterns and protein epitopes
Cells grown on almost any substrate are able to be treated with CellCover. This includes tissue samples destined for a biobank. Cells fixed with CellCover can be kept without worry of undesired changes due to storage conditions. Immediately after this unique type of fixation, such cells maintain near-perfect in vivo morphology — and without the need for traditional chemical crosslinking of biomolecules.
RNA degradation is also inhibited (as measured by high RIN values) leading to great results in RNA sequencing or single cell sequencing experiments. Even the integrity of high molecular weight RNA is protected (e.g precurser rRNA, which are normally degraded after applying commonly used standards procedures) – and there is no need to hurry! You can store your sample in CellCover at 4°C and isolate RNA later.
Remarkably, protein expression analysis appear to have no limits when using CellCover. In contrast to conventional formaldehyde fixation which mask formalin sensitive epitopes, these remain readily accessible for antibody binding in immunohistochemical experiments.
Treat now — analyze later and then analyze again!
A special benefit of CellCover is that you can perform additional molecular analyses on cells that have already been analyzed once on a morphological or molecular level. For example, DNA (e.g. PCR), RNA (e.g RNA sequencing), or protein (e.g protein modification analysis) can be isolated and analyzed, even after after initial analysis by IHC, ICC, or flow cytometry.
This unique property enables new possibilities for analyzing cellular functions. For example, after fixation with CellCover, subpopulations of cells can be separated according to known properties by flow cytometry and subsequently analyzed for gene expression profiles on RNA or on protein-level. This makes it is easy to link characteristics of the transcriptome and the proteome for a given cell population.
- Seed and grow cells on chamber slides
- Remove medium
- Wash cells 1x with PBS or CellCover
- Place slide in CellCover and store at 4°C until use
- Proceed to staining protocol according to experimental design, e.g. immunostaining (If RNA is to be isolated for downstream application, you can stain cells by using CellCover)
- Remove cell culture media
- Add appropriate volume of CellCover and incubate for 2-3 min at room temperature
- Remove CellCover
- Add trypsin solution (e.g. Trypsin/EDTA 0,25% for adherent cells) and incubate
for 5 min at 37°C (or until single cells detached)
- Collect cells by centrifugation
- Resuspend cells in CellCover and store at 4°C until needed or proceed with your
- Harvest cells by centrifugation
- Remove supernatant
- Flick tube to suspend cells in residual medium
- Add CellCover (5 to 10x volumes) and store at 4°C until use
- Pellet cells and remove SN
(Some cells might need higher centrifugation speed for pelleting.)
- Proceed to standard staining protocols, e.g. immunostaining (If RNA is to be isolated for downstream application, you can stain cells by using CellCover as antibody diluent and washing buffer
- Cut tissue into approximately 5 mm pieces
- CellCover does not penetrate tight junctions, thus encapsulated organs must be open across the largest diameter.
- Place tissue in CellCover (at least 10x volumes) and store at 4°C until use
- Change CellCover at least once after 4-24 hours
- Proceed according to experimental design e.g. RNA isolation
Stabilization of molecules in biological material
800-050; 800-125, 800-250, 900-196
|Sizes||50ml, 125ml, 250ml, 196 x 10ml|
|Incubation Time||2-5 minutes for cells
Overnight at 4°C for tissues
|Sample Storage Post-Treatment||RT – Up to 24 hours
4°C – Up to 12 days
|Cell Type Compatibility||Suspension, adherent, tissue|
|Storage||1 year at 4°C|