Product Name: HybriCare™ Hybridoma Rescue Media Supplement, 20x
Catalogue No.: 113003
Description: HybriCare™ Hybridoma Rescue Media Supplement, is a proprietary blend of growth factors, cytokines, hormones & antibiotics, when added to your current media at 5% will help rescue your clone from poor viability or productivity.
HybriCare™ is based on our HyMax™, our proprietary Hybridoma Supplement, as basal media. Hybridoma clones tend to lose viability or productivity spontaneously and randomly for various reasons. These includes loss of gene expression and non-obvious contamination such as mycoplasma. Using our extensive cell culture experience, we have specially formulated this media that contains many cytokines, growth factors and hormones which helps B-cells to retain their IgG isotype phenotype. It also contains some antibiotics to stop the invisible contamination such as mycoplasmales, mycotics & some bacteria. We are successful at rescuing over 80% of cell lines provided to us. It is prepared fresh and intended to be used within 6 months.
Format: Provided as a 20x concentrate ready to use solution. Never frozen before shipping to customer. Each lot is tissue culture tested.
Storage: It is recommended to store at 4°C - 8°C in refrigerator until use within 6 months. It
can be made into aliquots and frozen stored at -20°C - 80°C for long term to use within 12 months period. Avoid repeated freezing and thawing, which may cause activity loss and or show precipitation.
Stability: 6 months when stored at 4°C - 8°C in refrigerator or 12 months when stored frozen.
Once thawed use within 30 days or aliquot and store as needed. Do not freeze & thaw repeatedly.
Shipping: It is prepared fresh (within 30 days) and shipped in liquid form on ice packs.
Protocol and Instruction for Use
Notes: Read these instructions before use
1. Some precipitate may appear in HybriCare after shipment, or storage in refrigerator or upon frozen storage (it is not contamination). If so, warm the solution to 37°C for about half an hour, typically the precipitate is expected to go back into solution. Any remaining precipitate will go into solution once it is added to working media at 5%. Presence of light precipitate will not affect cell culture. Alternatively, it may also be sterile filtered before use.
2. Do not use HybriCare at more than 10% of your working media.
3. Recommended media are DMEM, RPMI, and MEM with or without other additives or serum.
4. Some microbial contamination is invisible to naked eye, with only signs of poor growth or viability of culture. Common antibiotics used in cell culture such as penicillin and streptomycin or gentamycin will not help. HybriCare also contains some selective antibiotics to rescue such cultures.
Method of use:
1. Add 5mL of HybriCare to 95mL of your media currently being used (including any supplements or antibiotics) for culturing cells to make the working media. Mix well.
2. Use the prepared HybriCare working media for culturing your cells as you would normally use.
3. Continue with culturing cells with this media until cells have grown well for few generations.
4. If microbial contamination is suspected in your culture, use HybriCare for at least 6 generations before weaning off. Once cells have grown well, culture may be weaned off of HybriCare by gradually reducing the percentage in steps to 1%, 0.5%, 0.1% and 0%.
• If mycoplasma contamination is suspected in your culture, we have MycoLytix 1000x, a ready to use mycoplasma inhibitor solution (Item 113015) to rescue such cultures.
• We also offer guaranteed cell line rescue as custom service. We do all the work for you and you pay only if we are able to rescue your clone.
• Check our website to see our complete list of cell culture and hybridoma reagents. Contact us for more information.