ClonaCell®-HY Medium A

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Pre-fusion Myeloma Growth Medium and Hybridoma Expansion Medium

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  • ClonaCell®-HY 500 mL
  • Label for ClonaCell®-HY 500 mL
ClonaCell®-HY 500 mL
ClonaCell®-HY Medium A supports the growth of myeloma cells prior to fusion and expansion of hybridomas post-selection. This medium is used during mouse monoclonal antibody development.
Product Name Description Catalog # Size Price Quantity
ClonaCell®-HY Medium A Pre-fusion myeloma growth medium and hybridoma expansion medium 03801 500 mL 109.00 USD      
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Recommended for:
Growth and expansion of parental myeloma cells prior to fusion and expansion of stable hybridoma clones
Intended Use Statement: For Research Use Only. Not for Therapeutic or Diagnostic Use.
Contains:
• DMEM
• Pre-selected serum
• Gentamycin
• Supplements
Product Type: Hybridoma generation & transfected cell selection
Application: Hybridoma generation, Cell line development
Area of Interest: Cell line development, Hybridoma generation, Pharmacology, toxicology, drug discovery
Cell Type: Hybridomas
Popular Product Line: ClonaCell

Procedures and instruction manuals:

Educational resources:

MSDS:

FAQS:


  • Q. WHY IS THERE HT (HYPOXANTHINE, THYMIDINE) IN MEDIUM E?

    A. Hybridomas are selected using HAT (Hypoxanthine, Aminopterin, Thymidine), which contains aminopterin. Aminopterin blocks the de novo pathway for synthesizing DNA, and the effect can persist even after the cells are removed from selection. HT provides the necessary substrates for the cells to synthesize DNA using the salvage pathway.

  • Q. IS THE SERUM IN CLONACELL® HEAT INACTIVATED?

    A. Yes, all serum used in ClonaCell® is heat inactivated.

  • Q. IS THERE ANY IG OR INSULIN IN THE CLONACELL® MEDIA?

    A. While we don't add Ig or insulin specifically to the ClonaCell® Media, we do add serum, which contains an undefined amount of immunoglobulins and insulin.

  • Q. HOW DO I THAW MEDIUM D (METHYLCELLULOSE)?

    A. We recommend thawing the medium overnight in a refrigerator at 4oC and mixing well.

  • Q. MY MEDIUM D (METHYLCELLULOSE) APPEARS RUNNY. WHY DOES THIS HAPPEN?

    A. “Runny” methylcellulose could be a result of improper handling.  Over-dilution of methylcellulose, aliquoting with a pipette rather than a syringe and blunt-end needle, or insufficient mixing before use will all result in methylcellulose having altered viscosity.  Note: Methylcellulose is less viscous at RT than 37oC.

  • Q. WHY DO I HAVE TO PUT MY FUSED CELLS INTO LIQUID MEDIUM OVERNIGHT? WHY CAN’T I JUST PLATE DIRECTLY INTO MEDIUM D?

    A. We recommend waiting up to 24 hours so that all of the fused cells can go through one cell cycle. This will ensure they have a chance to express HPRT (Hypoxanthine Guanine Phosphoribosyltransferase), the enzyme necessary to survive in the presence of aminopterin (present in Medium D). Additionally, fused cells are very fragile immediately after fusion. Waiting a day before mixing the cells with the methylcellulose will improve their survival.

  • Q. CAN I GROW HUMAN/RAT/T CELL HYBRIDOMAS IN CLONACELL®-HY?

    A. Although we have not tried to generate human, rat or T cell hybridomas, there is no reason they should not work in ClonaCell®-HY. The researcher needs to ensure that the cell lines used in the fusion are sensitive to HAT selection, and grow well in methylcellulose-based medium.

  • Q. DO I EVER NEED TO RECLONE HYBRIDOMAS GROWN WITH CLONACELL®-HY?

    A. Recloning may be necessary if the number of colonies in the original dishes was very high.

  • Q. THERE ARE VERY FEW COLONIES GROWING IN MY MEDIUM D. WHY?

    A. Low numbers of colonies is generally a result of low fusion efficiency, which can have many causes. The fusion efficiency can be affected by the presence of serum during fusion, the presence of mycoplasma, low viability of cells, overexposure to PEG (polyethylene glycol), or slow growing myeloma cells prior to fusion.

  • Q. ONCE I PLUCK THE COLONIES AND GROW THE CELLS IN PLATES, WILL THE RESIDUAL METHYLCELLULOSE INTERFERE WITH CHARACTERIZATION? FOR EXAMPLE, WILL I HAVE PROBLEM DOING AN ELISA?

    A. There will likely be some residual methylcellulose contamination when colonies are plucked and transferred to the 96-well plate with the Medium E. However, the concentration of methylcellulose should be low enough that it should not show up as background.

Product Name

Description

Catalog #
ClonaCell®-HY Hybridoma Kit Complete Kit for Mouse Monoclonal Antibody Generation 03800
ClonaCell®-HY Medium B Hybridoma Fusion Medium 03802
ClonaCell®-HY Medium C Fusion Recovery Medium 03803
ClonaCell®-HY Medium D Methylcellulose-Based Hybridoma Selection and Cloning Medium 03804
ClonaCell®-HY Medium E Hybridoma Growth Medium Containing HT 03805
ClonaCell®-HY PEG Pre-tested Polyethylene Glycol for Cell Fusion 03806
Blunt-End Needles, 16 Gauge 16 Gauge Blunt-end Needles 28110
Blunt-End Needles 28120
100 mm Treated Tissue Culture Dishes 100 mm Treated Tissue Culture Dishes 27125
100 mm Treated Tissue Culture Dishes 27127
ClonaCell® EasyPick Robotic Colony Picking and Liquid Handling Instrument 30000

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