CLS-54 genomic DNA - 5 microgram
The CLS-54 cell was established from the primary lung carcinoma of a 65-year-old man in 1998.
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Biosafety level: |
1.. |
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Citation: |
CLS-54 (CLS catalog number 300227). |
General characteristics: |
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Organism: |
Human. |
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Ethnicity: |
Caucasian. |
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Age: |
65. |
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Gender: |
Male. |
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Tissue: |
Lung. |
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Disease |
Adenocarcinoma |
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Morphology |
Epithelial-like |
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Growth properties |
Adherent |
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Depositor |
H. Löhrke |
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Cellosaurus Accession |
CVCL_5728 |
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NCBI_TaxID |
9606 |
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Description |
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Culture conditions |
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Culture Medium |
RPMI 1640, 10% FBS |
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Medium supplements |
2 mM L-glutamine |
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Subculturing |
Remove medium and rinse the adherent cells using PBS without calcium and magnesium (3-5 ml PBS for T25, 5-10ml for T75 cell culture flasks). Add Accutase (1-2ml per T25, 2.5ml per T75 cell culture flask), the cell sheet must be covered completely. Incubate at ambient temperature for 8-10 minutes. Carefully resuspend the cells with medium (10 ml), centrifuge for 3 min at 300xg, resuspend cells in fresh medium and dispense into new flasks which contain fresh medium. |
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Split ratio |
A ratio of 1:2 to 1:6 is recommended |
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Seeding density |
1 x 10^4 cells/cm^2 will yield in a confluent layer in about 4 days |
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Fluid renewal |
Every 3 to 5 days |
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Freeze medium |
CM-ACF (CLS catalog number 800650) |
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Freezing recovery |
After thawing, plate the cells at 5 x 10^4 cells/cm^2 and allow the cells to recover from the freezing process and to adhere for at least 24 hours. |
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Sterility |
Plasmotest: negative; Mycoplasma specific PCR: negative |
Special features |
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Tumorigenic |
Yes, in nude mice |
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Viruses |
SMRV: Negative, as confirmed by Real-Time PCR |
STR profile |
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Amelogenin |
X,X |
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CSF1PO |
12 |
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D13S317 |
11 |
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D16S539 |
12,13 |
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D5S818 |
13 |
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D7S820 |
10,11 |
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TH01 |
6,9.3 |
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TPOX |
8,9 |
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vWA |
14,17 |
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D3S1358 |
18 |
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D21S11 |
30,31.2 |
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D18S51 |
11,17,18 |
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Penta E |
12,15 |
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Penta D |
9 |
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D8S1179 |
11 |
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FGA |
20 |
Handling of cells |
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Cryopreserved cells |
The cells come deep-frozen shipped on dry ice. Please make sure that the vial is still frozen. If immediate culturing is not intended, the cryovial must be stored below -150°C after arrival. If immediate culturing is intended, please follow the below instructions: Quickly thaw by rapid agitation in a 37 °C water bath within 40-60 seconds. The water bath should have clean water containing an antimicrobial agent. As soon as the sample has thawed, remove the cryovial from the water bath. A small ice clump should still remain and the vial should still be cold. From now on, all operations should be carried out under aseptic conditions. Transfer the cryovial to a sterile flow cabinet and wipe with 70% alcohol. Carefully open the vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of culture medium (room temperature). Resuspend the cells carefully. Centrifuge at 300 x g for 3 min and discard the supernatant. The centrifugation step may be omitted, but in this case the remains of the freeze medium have to be removed 24 hours later. Resuspend the cells carefully in 10 ml fresh cell culture medium and transfer them into two T25 cell culture flasks. All further steps are described in the Subculture section. |
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Proliferating Cultures |
One or two cell culture flasks come filled with cell culture medium. Collect the entire medium in 1 or 2 x 50 ml centrifuge tubes, respectively. Carefully add 5 ml of cell culture medium to each T25 cell culture flask. Control the cell morphology and confluency under the microscope. Incubate at 37 °C for a minimum of 24 hours. Spin down the collected medium at 300 x g for 3 minutes to collect the cells which may have detached during transit. If a cell pellet is visible, resuspend the cells in 5 ml of cell culture medium and transfer to a T25 cell culture flask. Incubate at 37 °C for a minimum of 24 hours. |
Documentation |
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Safety precautions |
If the cryovial is planned to be stored in liquid nitrogen and to be thawed in the future, special safety precautions should be followed. Protective gloves and clothing should be used and a facemask or safety goggles must be worn when transferring frozen samples into or removing from the liquid nitrogen tank. The removal of a cryovial from liquid nitrogen may result in the explosion of the frozen vial creating flying fragments. |
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Warranty |
CLS warrants for a high cell viability and culture performance only if the products are stored and cultured according to the information described in this product sheet. |
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Certificate of Analysis |
The certificate of analysis can be requested on the website or via email at info@cls.shop. Please indicate the lot number of your product in the email. |
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Disclaimer |
This product is intended for laboratory research use only. It is strictly prohibited to use the products in human or animal subjects for therapeutic, diagnostic or prophylactic purposes. |
Required products
In biological research, the cryopreservation of mammalian cells is an invaluable tool. Successful preservation of cells is a top priority given that losing a cell line to contamination or improper storage conditions leads to lost time and money, ultimately delaying research results. Once the cells have been transferred from a cell growth medium to a freezing medium, the cells are typically frozen at a regulated rate and stored in liquid nitrogen vapor or at below -130°C in a mechanical deep freezer. The freeze medium CM-ACF enables cryopreservation of cells at below -130°C (or in liquid nitrogen), essentially eliminating the need for an additional, costly ultralow freezer and eliminating time-consuming and demanding controlled rate freezing processes. Simply collect the cells, aspirate the growth medium, resuspend in CM-ACF, transfer to a cryovial, and store the vial at below -130 °C.
Long shelf-life
CM-ACF is a serum-free, ready-to-use cryopreservation medium that can be stored in the refrigerator for up to one year.
Trusted by hundreds of researchers
Our advanced, serum-free cell freezing medium CM-ACF is a market-leading product in Germany and Europe and is distinguished by numerous publications involving hundreds of different cell lines worldwide. We tested it with more than 1000 cell lines from our proprietary cell bank.
Optimized serum-free ingredients
CM-ACF does not contain serum products. Serum-containing cryopreservation mediums have the disadvantage of fluctuating recovery rates and unclear composition. Since the composition and concentration of proteins and other biological components vary from batch to batch in serum, the reproducibility of experiments with cells that were frozen in a serum-containing medium may be compromised. As each component of CM-ACF is carefully defined, you can rest assured that cells always recover identically.
Contains DMSO, glucose, salts
Buffering capacity pH = 7.2 to 7.6
Universal
- even for stem cell preservation
All common cell lines can be frozen and thawed to yield many viable cells. Compared to standard media, the rate of recovery of even the most delicate cells is significantly higher. Using CM-ACF, we store over 1000 different cell lines with outstanding success.
Applications & Validation
The cells preserved in our CM-ACF freeze medium can be used for cell counting, viability and cryopreservation, cell culture, mammalian cell culture, gene expression analysis and genotyping, in vitro transcription, and polymerase chain reactions. Each batch's efficacy is evaluated using CHO-K1 cells. Each batch is tested for pH, osmolality, sterility, and endotoxins to ensure high quality.
In biological research, the cryopreservation of mammalian cells is an invaluable tool. Successful preservation of cells is a top priority given that losing a cell line to contamination or improper storage conditions leads to lost time and money, ultimately delaying research results. Once the cells have been transferred from a cell growth medium to a freezing medium, the cells are typically frozen at a regulated rate and stored in liquid nitrogen vapor or at below -130°C in a mechanical deep freezer. The freeze medium CM-1 enables cryopreservation of cells at below -130°C (or in liquid nitrogen), essentially eliminating the need for an additional, costly ultralow freezer and eliminating time-consuming and demanding controlled rate freezing processes. Simply collect the cells, aspirate the growth medium, resuspend in CM-1, transfer to a cryovial, and store the vial at below -130 °C.
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.
Trusted by hundreds of researchers
Our advanced cell freezing medium CM-1 is a market-leading product in Germany and Europe and is distinguished by numerous publications involving hundreds of different cell lines worldwide. We tested it with more than 1000 cell lines from our proprietary cell bank.
Optimized serum-free ingredients
CM-1 does contain serum products. Serum-containing cryopreservation mediums optimally protect the cells whilst being frozen and have the advantage of high recovery rates. As CM-1 has been tested with a multitude of cell lines, you can rest assured that your cells always recover well.
Contains FBS, DMSO, glucose, salts
Buffering capacity pH = 7.2 to 7.6
Applications & Validation
The cells preserved in our CM-1 freeze medium can be used for cell counting, viability and cryopreservation, cell culture, mammalian cell culture, gene expression analysis and genotyping, in vitro transcription, and polymerase chain reactions. Each batch's efficacy is evaluated using CHO-K1 cells. Each batch is tested for pH, osmolality, sterility, and endotoxins to ensure high quality.
It is suitable for a range of applications, such as culturing primary cells, stem cells, and immortalized cell lines. RPMI 1640 is also used in various immunological assays, such as ELISAs and co-cultures. The RPMI-1640 medium has been validated extensively over the years to ensure its reliability and consistency. It is recommended by experts in the field of cell biology for use in experiments requiring robust growth and proliferation of mammalian cell types. Its composition makes RPMI 1640 an ideal choice for many complex research applications.
References
1) Merskey, R., & Pappenheimer, A. M. (1963). A New Medium For The Cultivation Of Leukemia Cells And Other Tissue-Culture Cells. Proceedings of the Society for Experimental Biology and Medicine, 112(2), 845–849.
2) RPMI-1640: a protein-free medium optimized for growth characteristics of human hematopoietic cells. (1984). In Vitro Cellular & Developmental Biology, 20(4), 467–475.
- 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) 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.
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