B-LCL-HROC24 Cells
General information
Organism | Human |
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Tissue | Peripheral blood |
Disease | Carcinoma |
Synonyms | B-LCL CO24, Bc HROC24 |
Characteristics
Age | 98 years |
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Gender | Male |
Ethnicity | Caucasian |
Morphology | Round cells |
Cell type | B lymphoblast |
Growth properties | Suspension |
Identifiers / Biosafety / Citation
Citation | B-LCL-HROC24 (Cytion catalog number 300861) |
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Biosafety level | 2 |
Depositor | M. Linnebacher |
Expression / Mutation
Surface antigens | CD19 |
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Handling
Culture Medium | RPMI 1640, w: 2.1 mM stable Glutamine, w: 2.0 g/L NaHCO3 (Cytion article number 820700a) |
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Medium supplements | Supplement the medium with 10% heat-inactivated FBS |
Subculturing | Gently homogenize the cell suspension in the flask by pipetting up and down, then take a representative sample to determine the cell density per ml. Dilute the suspension to achieve a cell concentration of 1 x 10^5 cells/ml with fresh culture medium, and aliquot the adjusted suspension into new flasks for further cultivation. |
Freeze medium | CM-1 (Cytion catalog number 800100) or CM-ACF (Cytion catalog number 806100) |
Handling of cryopreserved cultures |
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Quality control / Genetic profile / HLA
Sterility | Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections. |
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STR profile |
Amelogenin: x,y
CSF1PO: 11,12
D13S317: 7,13
D16S539: 9,11
D5S818: 10,11
D7S820: 11,13
TH01: 6,9
TPOX: 11
vWA: 16,2
D21S11: 27,28
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HLA alleles |
A*: 02:01:01, 30:02:01
B*: 18:01:01, 38:01:01
C*: 05:01:01, 12:03:01
DRB1*: 03:01:01, 13:01:01
DQA1*: 01:03:01, 05:01:01
DQB1*: 02:01:01, 06:03:01
DPB1*: 02:02:01, 03:01:01
E: 01:01:01, 01:03:02
|
Required products
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.
This unique formulation combines Dulbecco's Modified Eagle Medium (DMEM) and Ham's F-12 (Ham's Nutrient Mixture F-12) in a precise 1:1 ratio. The addition of L-glutamine further enhances its composition.
DMEM, derived from Eagle's Minimal Essential Medium (EMEM), offers an increased concentration of amino acids and vitamins compared to its predecessor. In contrast, Ham's F-12 is based on Ham's F-10 medium, providing a complementary set of essential components.
To support optimal cell growth, it is common practice to supplement DMEM:Ham's F12 with FBS at a typical concentration of 5-10%. This addition is necessary as the medium lacks growth hormones, lipids, and proteins crucial for cellular development.
DMEM:Ham's F12 incorporates a pH buffer system and is often supplemented with phenol red, a pH indicator. Cultured cells in DMEM:Ham's F12, or any medium utilizing the bicarbonate buffer system, require a controlled CO2 environment of 5-10% to maintain appropriate pH levels. Phenol red enables monitoring of pH changes from 6.2 (yellow) to 8.2 (red).
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 as well as 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
154,45
Iron(III)-nitrate x 9H2O
0,05
Iron(II)-sulfate x 7H2O
0,42
Potassium chloride
311,83
Copper(II)-sulfate x 5H2O
0.001
Magnesium chloride anhydrous
28,57
Magnesium sulfate
48,85
Sodium chloride
6,999.50
Sodium dihydrogen phosphate anhydrous
54,35
di-Sodium hydrogen phosphate
70,98
Zinc sulfate x 7H2O
0,43
Other Components
D(+)-Glucose anhydrous
3,151.00
HEPES
3,574.50
Hypoxanthine
2,04
Linoleic acid
0,04
DL-68-Lipoic acid
0.103
Sodium pyruvate
110,00
Phenol red
8,10
Putrescin x 2HCl
0.081
Thymidine
0,36
Amino Acids
L-Alanine
4,45
L-Arginine x HCl
147,35
L-Asparagine x H2O
7,50
L-Aspartic acid
6,65
L-Cystine x 2HCl
31,29
L-Cysteine x HCl x H2O
17,56
L-Glutamine
365,00
L-Glutamic acid
7,35
Glycine
18,75
L-Histidine x HCl x H2O
31,48
L-Isoleucine
54,37
L-Leucine
58,96
L-Lysine x HCl
91,37
L-Methionine
17,24
L-Phenylalanine
35,48
L-Proline
17,27
L-Serine
26,25
L-Threonine
53,55
L-Tryptophan
9,02
L-Tyrosine x 2Na x 2H2O
55,81
L-Valine
52,66
Vitamins
D-(+)-Biotine
0,004
D-Calcium pantothenate
2,12
Choline chloride
8,98
Folic acid
2,66
myo-Inositol
12,51
Nicotinamide
2,02
Pyridoxine x HCl
2,03
Riboflavin
0,22
Thiamine x HCl
2,17
Vitamine B12
0,68
NaHCO3
1,200.00
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