ClonaCell™-HY AOF Expansion Medium
Hybridoma expansion medium with hypoxanthine and thymidine (animal origin-free)
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ClonaCell™-HY AOF Expansion Medium -
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Overview
ClonaCell™-HY AOF Expansion Medium is an animal origin-free (AOF) and serum-free liquid medium optimized for hybridoma expansion after hypoxanthine, aminopterin, thymidine (HAT) selection. The medium contains hypoxanthine and thymidine (HT) and is used to wean hybridomas off aminopterin used during the selection process. ClonaCell™-HY AOF Expansion Medium promotes robust expansion of hybridomas, adaptation from serum-containing to serum-free media, and stabilizing the viability and antibody productivity of hybridoma cell lines. The majority of hybridomas can be switched directly from serum-containing media to ClonaCell™-HY AOF Expansion Medium without an adaptation step. The absence of serum in this medium facilitates detection and purification of the desired hybridoma-derived antibody without interference from serum-derived immunoglobulins.
This medium has been verified for use with mouse and rat hybridomas and is suitable for expansion of mouse myelomas such as Sp2/0, rat myelomas such as YB2/0, and human myelomas such as U266. In most cases, cryopreserved hybridomas can be thawed directly into ClonaCell™-HY AOF Expansion Medium while maintaining a high level of viability. This medium may be used as a serum-free alternative to ClonaCell™-HY Medium A (Catalog #03801) or ClonaCell™-HY Medium E (Catalog #03805). No materials of animal or human origin are used in the manufacture of this medium or its components, to at least the secondary level of manufacturing.
This medium has been verified for use with mouse and rat hybridomas and is suitable for expansion of mouse myelomas such as Sp2/0, rat myelomas such as YB2/0, and human myelomas such as U266. In most cases, cryopreserved hybridomas can be thawed directly into ClonaCell™-HY AOF Expansion Medium while maintaining a high level of viability. This medium may be used as a serum-free alternative to ClonaCell™-HY Medium A (Catalog #03801) or ClonaCell™-HY Medium E (Catalog #03805). No materials of animal or human origin are used in the manufacture of this medium or its components, to at least the secondary level of manufacturing.
Advantages:
• Animal origin-free
• Absence of serum reduces performance variability in the medium
• Simplifies downstream clone screening and antibody purification (no serum-derived IgG)
• Absence of serum reduces performance variability in the medium
• Simplifies downstream clone screening and antibody purification (no serum-derived IgG)
Contains:
• Hypoxanthine and thymidine (HT)
• Gentamicin
• Phenol red
• L-Glutamine and other supplements
• Other ingredients, including recombinant proteins
• Gentamicin
• Phenol red
• L-Glutamine and other supplements
• Other ingredients, including recombinant proteins
Subtype:
Specialized Media
Cell Type:
Hybridomas
Species:
Human; Mouse; Rat
Application:
Cell Culture
Brand:
ClonaCell
Area of Interest:
Antibody Development; Cell Line Development; Hybridoma Generation
Formulation:
Serum-Free
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Complete kit for hybridoma generation -
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Hybridoma growth medium with hypoxanthine and thymidine (serum-containing)
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Figure 1. Cloning Efficiencies for Three Hybridoma Cell Lines Subcloned in Serum-containing and Serum-free Cell Culture Media
The hybridoma lines were adapted to growth in each medium and subcloned by limiting dilution (n = 1 - 5). After incubation for 10 days (37°C, 5% CO2), the plates were analyzed with a Cell Metric™ instrument (Solentim). The plating efficiency was estimated by Poisson statistics using the ELDA method described by Hu & Smith, 2009 (J Immunol Meth, 347: 70-78). ACF = animal component-free. Data is expressed as mean + 1 SD.
Figure 2. Expansion of an Established Hybridoma Cell Line in Several Commercially-Available Serum-Containing and Serum-Free Hybridoma Cell Culture Media
The hybridoma line was adapted to growth in 13 different cell culture media: ClonaCellTM-HY AOF Expansion Medium (animal origin-free), DMEM + 10% FBS or ClonaCellTM-HY Medium E (serum-containing), Medium 1 (serum-free with serum-derived proteins), Medium 2 – 4 (serum-free), Medium 5 (animal component-free), Medium 7, 9 (protein-free), and Medium 11 – 13 (chemically-defined). They were then seeded in triplicate in 24-well tissue culture plates at 5 x 103 cells/mL. The cultures were incubated at 37°C (5% CO2 atmosphere) and the viable cell density was measured on days 3 – 7. Data is expressed as mean ± 1SD.
PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.