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ClonaCellâ„¢-HY AOF Expansion Medium

Hybridoma expansion medium with hypoxanthine and thymidine (animal origin-free)

ClonaCellâ„¢-HY AOF Expansion Medium

Hybridoma expansion medium with hypoxanthine and thymidine (animal origin-free)

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Hybridoma expansion medium with hypoxanthine and thymidine (animal origin-free)
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Product Advantages


  • Animal origin-free

  • Absence of serum reduces performance variability in the medium

  • Simplifies downstream clone screening and antibody purification (no serum-derived IgG)

Overview

ClonaCellâ„¢-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. ClonaCellâ„¢-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 ClonaCellâ„¢-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 ClonaCellâ„¢-HY AOF Expansion Medium while maintaining a high level of viability. This medium may be used as a serum-free alternative to ClonaCellâ„¢-HY Medium A (Catalog #03801) or ClonaCellâ„¢-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.
Contains
• Hypoxanthine and thymidine (HT)
• 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 Category
Serum-Free

Data Figures

Cloning Efficiencies for Three Hybridoma Cell Lines Subcloned in Serum-containing and Serun-free Cell Culture Media

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.

Graph comparing growth of hybridoma cell line expanded in commercially available serum-containing and serum-free hybridoma cell culture media.

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.

Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

Document Type
Product Name
Catalog #
Lot #
Language
Document Type
Product Name
Catalog #
03835
Lot #
All
Language
English
Document Type
Product Name
Catalog #
03835
Lot #
All
Language
English
Document Type
Product Name
Catalog #
03835
Lot #
All
Language
English

Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

Resources and Publications

Publications (1)

MUC13 negatively regulates tight junction proteins and intestinal epithelial barrier integrity via protein kinase C C. Segui-Perez et al. Journal of Cell Science 2024 Mar

Abstract

Glycosylated mucin proteins contribute to the essential barrier function of the intestinal epithelium. The transmembrane mucin MUC13 is an abundant intestinal glycoprotein with important functions for mucosal maintenance that are not yet completely understood. We demonstrate that in human intestinal epithelial monolayers, MUC13 localized to both the apical surface and the tight junction (TJ) region on the lateral membrane. MUC13 deletion resulted in increased transepithelial resistance (TEER) and reduced translocation of small solutes. TEER buildup in ΔMUC13 cells could be prevented by addition of MLCK, ROCK or protein kinase C (PKC) inhibitors. The levels of TJ proteins including claudins and occludin were highly increased in membrane fractions of MUC13 knockout cells. Removal of the MUC13 cytoplasmic tail (CT) also altered TJ composition but did not affect TEER. The increased buildup of TJ complexes in ΔMUC13 and MUC13-ΔCT cells was dependent on PKC. The responsible PKC member might be PKCδ (or PRKCD) based on elevated protein levels in the absence of full-length MUC13. Our results demonstrate for the first time that a mucin protein can negatively regulate TJ function and stimulate intestinal barrier permeability.