Murine leukemia virus

Infect cells when cultures are approximately 60% confluent. Treat with DEAE dextran (25 µg/mL) or Polybrene (8 µg/mL) for 1 hour. Wash the cells with medium or PBS. Inoculate the cell monolayer when still sub-confluent and feed with complete medium. One hour adsorption is recommended. The next day the monolayer is almost confluent. Change the medium at the end of the day. Harvest the supernatant the next morning. The virus is very labile and does not survive long at 37°C. Filter through a 45U filter and quick freeze. Refeed the cells with complete medium and repeat the harvest the next morning. Proceed until the cells eventually slough off or enough supernatant has been harvested. The virus is cell associated, therefore at the end of the harvest, the cells can be scraped in 3-4 mL medium, frozen and thawed once and spun at 2500 RPM for 20 minutes and the supernatant added to the pool to increase titer. Pool the harvests, aliquot and quick freeze. Note: One T150 flask will produce approximately 100 mL supernatant. Titration has to be done by XC plaque assay on either NIH3T3 or SC-1 cells.

If the inoculum is persistently infected cells, grow the cells and split 1:3 until enough cells are produced. Slow freeze the cells. For titration, use the supernatant from the monolayer. Change the medium the day before , harvest the supernatant and run the XC plaque assay.

The product is provided 'AS IS' and the viability of ATCC^® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.  Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.

This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.