U2OS-CRISPR-NUP96-mEGFP Cells

McCoy's 5A Medium is a highly recommended and specialized medium designed to facilitate the growth and replication of viruses in primary cell cultures. It has gained significant recognition for its exceptional performance in various biological research applications.
One prominent application of McCoy's 5A Medium is its utilization in the culturing of human colon carcinoma cell lines. Specifically, it has been employed in the study of the leucine-rich repeat-containing G-protein-coupled receptor (LGR5) and its role in the metastasis of colon cancer. This medium has been effectively employed in the cultivation of several colon carcinoma cell lines, including HCT116, RKO, FET, CBS, HCT116b, and TENN, enabling researchers to delve deeper into the mechanisms underlying colon cancer metastasis.
In addition to its application in cancer research, McCoy's 5A Medium has proven to be indispensable in the study of osteoblasts. Researchers investigating the ion reactivity of calcium-deficient hydroxyapatite in standard cell culture media have utilized this medium to culture osteoblasts. This application has facilitated a better understanding of the interactions between osteoblasts and calcium-deficient hydroxyapatite, contributing to advancements in the field of bone research.
Notably, McCoy's 5A Medium was meticulously formulated by modifying the amino acids found in Basal Medium Eagle to provide optimal support for liver tumor cells. This enriched formulation enables its suitability for a diverse range of established cell lines, as well as primary cells, further enhancing its versatility and applicability in various research settings.
Moreover, McCoy's 5A Medium extends its biochemical and physiological effects beyond liver tumor cells. It has been successfully employed to support growth in primary cultures of bone marrow, skin, gingiva, kidney, omentum, adrenal, lung, spleen, rat embryo, and other cell types. This wide range of applications attests to the broad utility of McCoy's 5A Medium in supporting the growth and maintenance of various cell types for comprehensive biological research.
Formulation
This McCoy's 5A medium (modified) contains 3.0 g/L of Glucose, stable Glutamine, 2.0 mM of Sodium pyruvate, and 2.2 g/L of NaHCO3.
Quality control

pH = 7.2 +/
- 0.02 at 20-25°C. 
Each lot has been tested for sterility and absence of mycoplasma and bacteria.

Maintenance

Keep refrigerated at +2°C to +8°C in the dark. Freezing and warming up to +37° C minimize the quality of the product. 
Do not heat the medium to more than 37° C or use uncontrollable sources of heat (e.g., microwave appliances). 
If only a part of the medium is to be used, remove this amount from the bottle and warm it up at room temperature.
Shelf life for any medium except for the basic medium is 8 weeks from the date of manufacture.
Composition




Components
mg/L


Inorganic Salts
Calcium chloride x 2H2O
132,46



Magnesium sulfate x 7H2O
200,00



Potassium chloride
400,00



Sodium chloride
6,460.00



Sodium dihydrogen phosphate x H2O
580,00


Other Components
D(+)-Galactose anhydrous
3,000.00



Glutathione (red.)
0,50



Bacto-Peptone
600,00



Phenol red
10,00


Amino Acids
L-Alanine
13,36



L-Arginine x HCl
42,10



L-Asparagine x H2O
45,00



L-Aspartic acid
19,97



L-Cysteine
24,24



L-Glutamine stable
326,61



L-Glutamic acid
22,10



Glycine
7,50



L-Histidine x HCl x H2O
20,76



L-Hydroxyproline
19,70



L-Isoleucine
39,36



L-Leucine
39,36



L-Lysine x HCl
36,50



L-Methionine
14,90



L-Phenylalanine
16,50



L-Proline
17,30



L-Serine
26,30



L-Threonine
17,90



L-Tryptophan
3,10



L-Tyrosine
18,10



L-Valine
17,60


Vitamins
p-Aminobenzoic acid
1,00



Ascorbic acid
0,50



D(+)-Biotin
0,20



D-Calcium pantothenate
0,20



Choline chloride
5,00

Cytion’s Freeze Medium CM-1 is a state-of-the-art cryopreservation medium designed to ensure the highest level of cell viability and functionality post-thaw. This versatile medium is suitable for a broad spectrum of cell types, including both human and animal cells, making it an essential tool for diverse research applications. Formulated with a meticulously balanced combination of cryoprotectants and essential nutrients, Freeze Medium CM-1 minimizes ice crystal formation and cellular stress during the freezing process, thus preserving cellular integrity.
Key features of Freeze Medium CM-1 include:

Broad Compatibility: Effective for a wide range of cell types, including primary cells, stem cells, and established cell lines.
High Viability: Optimized to maximize post-thaw cell recovery and viability, ensuring reliable experimental outcomes.
Ready-to-Use: Conveniently prepared and sterilized for immediate application, reducing preparation time and risk of contamination.
Enhanced Stability: Maintains consistent performance under standard cryopreservation conditions, ensuring reproducible results.
Long Shelf Life: CM-1 is a serum-containing, ready-to-use cryopreservation medium that can be stored in the refrigerator for up to one year.


Using CM-1 for Freezing Cells

To use CM-1 for freezing both adherent and suspension cells, follow these steps:


For adherent cells, wash and dissociate them from the culture substrate. For suspension cells, proceed directly to the next step.
Count the cells to ensure they are at the proper concentration.
Centrifuge the cells to pellet them, then resuspend in CM-1 freeze medium.
Transfer the resuspended cells into cryovials.
Use a slow-freezing method before transferring the cells to long-term storage.





🥶 Method
🔍 Description
💡 Steps





❄️
Manual Freezing


A step-by-step method involving gradual temperature reduction to ensure cell viability.


1️⃣ Place cells in freeze medium in a 4°C freezer for 40 minutes.
2️⃣ Transfer to a -80°C freezer for 24 hours.
3️⃣ Store cells in liquid nitrogen for long-term preservation.




🧊
Using Mr. Frosty


A convenient device that allows for controlled freezing rates without electrical power.


1️⃣ Prepare cells in cryovials with freeze medium.
2️⃣ Place cryovials in Mr. Frosty container.
3️⃣ Store at -80°C for 24 hours before transferring to liquid nitrogen.




🧬
Controlled-Rate Freezer


A high-precision freezer by Thermo Fisher or other manufacturers designed for controlled temperature reduction.


1️⃣ Program the device to gradually decrease the temperature.
2️⃣ Place prepared cells in the freezer.
3️⃣ After the freezing cycle, transfer cells to liquid nitrogen.










Store the cryovials at temperatures below -130°C or in liquid nitrogen for long-term preservation.


Ingredients

Contains FBS, DMSO, Glucose, Salts
Buffering capacity: pH = 7.2 to 7.6


Cytion’s Freeze Medium CM-1 offers a reliable solution for cryopreservation, ensuring high cell viability and functionality post-thaw for a wide range of research applications.

Accutase Cell Dissociation Reagent
- A Gentle Alternative to Trypsin
Accutase is a cell detachment solution that is revolutionizing the cell culture industry. It is a mix of proteolytic and collagenolytic enzymes that mimics the action of trypsin and collagenase. Unlike trypsin, Accutase does not contain any mammalian or bacterial components and is much gentler on cells, making it an ideal solution for the routine detachment of cells from standard tissue culture plasticware and adhesion coated plasticware. In this blog post, we will explore the benefits and uses of Accutase and how it is changing the game in cell culture.
Advantages of Accutase
Accutase has several advantages over traditional trypsin solutions. Firstly, it can be used whenever gentle and efficient detachment of any adherent cell line is needed, making it a direct replacement for trypsin. Secondly, Accutase works extremely well on embryonic and neuronal stem cells, and it has been shown to maintain the viability of these cells after passaging. Thirdly, Accutase preserves most epitopes for subsequent flow cytometry analysis, making it ideal for cell surface marker analysis.
Additionally, Accutase does not need to be neutralized when passaging adherent cells. The addition of more media after the cells are split dilutes Accutase so it is no longer able to detach cells. This eliminates the need for an inactivation step and saves time for cell culture technicians. Finally, Accutase does not need to be aliquoted, and a bottle is stable in the refrigerator for 2 months.
Applications of Accutase
Accutase is a direct replacement for trypsin solution and can be used for the passaging of cell lines. Additionally, Accutase performs well when detaching cells for the analysis of many cell surface markers using flow cytometry and for cell sorting. Other downstream applications of Accutase treatment include analysis of cell surface markers, virus growth assay, cell proliferation, tumor cell migration assays, routine cell passage, production scale-up (bioreactor), and flow cytometry.
Composition of Accutase
Accutase contains no mammalian or bacterial components and is a natural enzyme mixture with proteolytic and collagenolytic enzyme activity. It is formulated at a much lower concentration than trypsin and collagenase, making it less toxic and gentler, but just as effective.
Efficiency of Accutase
Accutase has been shown to be efficient in detaching primary and stem cells and maintaining high cell viability compared to animal origin enzymes such as trypsin. 100% of cells are recovered after 10 minutes, and there is no harm in leaving cells in Accutase for up to 45 minutes, thanks to autodigestion of Accutase.
In summary
In conclusion, Accutase is a powerful solution that is changing the game in cell culture. With its gentle nature, efficiency, and versatility, Accutase is the ideal alternative to trypsin. If you are looking for a reliable and efficient solution for cell detachment, Accutase is the solution for you.

Phosphate-Buffered Saline (PBS) Solution: The Optimal Buffer for Your Biological Research
Phosphate-buffered saline (PBS) is a versatile buffer solution used in many biological and chemical applications, as well as tissue processing. Our PBS solution is formulated with high-quality ingredients to ensure a constant pH during experiments. The osmolarity and ion concentrations of our PBS solution are matched to those of the human body, making it isotonic and non-toxic to most cells.
Composition of our PBS Solution
Our PBS solution is a pH-adjusted blend of ultrapure-grade phosphate buffers and saline solutions. At a 1X working concentration, it contains 137 mM NaCl, 2.7 mM KCl, 8 mM Na2HPO4, and 2 mM KH2PO4. We have chosen this composition based on CSHL protocols and Molecular cloning by Sambrook, which are well-established standards in the research community.
Applications of our PBS Solution
Our PBS solution is ideal for a wide range of applications in biological research. Its isotonic and non-toxic properties make it perfect for substance dilution and cell container rinsing. Our PBS solution with EDTA can also be used to disengage attached and clumped cells. However, it is important to note that divalent metals such as zinc cannot be added to PBS as this may result in precipitation. In such cases, Good's buffers are recommended. Moreover, our PBS solution has been shown to be an acceptable alternative to viral transport medium for the transport and storage of RNA viruses, such as SARS-CoV-2.
Storage of our PBS Solution
Our PBS solution can be stored at room temperature, making it easy to use and access.
To sum up
In summary, our PBS solution is an essential component in many biological and chemical experiments. Its isotonic and non-toxic properties make it suitable for numerous applications, from cell culture to viral transport medium. By choosing our high-quality PBS solution, researchers can optimize their experiments and ensure accurate and reliable results.
Composition




Components
mg/L


Inorganic Salts
Potassium chloride
200,00



Potassium dihydrogen phosphate
200,00



Sodium chloride
8,000.00



di-Sodium hydrogen phosphate anhydrous
1,150.00