BM-N

1. Check all containers for leakage or breakage.
2. Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below ­-130°C, preferably in liquid nitrogen vapor, until ready for use.

• 2.6 g/L Tryptose Phosphate Broth (TPB)
• 0.069 mg/L ZnSO₄·7H₂O
• 3.86 mg/L AlCl₃·6H₂O
• 5 mg/L chlorophenol red
• 2g/L NaCl
• 10% heat-inactivated insect grade fetal bovine serum

Adjust pH to 6.3 to 6.4 by adding 1N NaOH
Adjust osmolarity to 317 to 360 by adding extra NaCl

The IPL-41 medium formulation was devised for use in a free gas exchange with atmospheric air. A CO₂ and air mixture is detrimental to cells when using this medium for cultivation. (In Vitro 17: 495-509, 1981)

Handling Procedure for Frozen Cells

To insure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C.  Storage at -70°C will result in loss of viability.

SAFETY PRECAUTION: ATCC highly recommends that protective gloves and clothing always be used and a full face mask always be worn when handling frozen vials.  It is important to note that some vials leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen.  Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vessel exploding or blowing off its cap with dangerous force creating flying debris.

1.     Thaw the vial by gentle agitation in a 27°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water.  Thawing should be rapid (approximately 2 minutes).

2.     Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.

3.     Transfer the vial contents to a centrifuge tube containing 9.0 ml complete growth medium and spin at approximately 125 x g for 5 to 7 minutes.

4.      Discard the supernatant and resuspend the cell pellet in an appropriate amount of fresh growth medium.

5.     Transfer the vial contents to an appropriate size vessel. 

Incubate the culture at 27°C in a suitable incubator. The IPL-41 medium formulation was devised for use in a free gas exchange with atmospheric air. A CO₂ and air mixture is detrimental to cells when using this medium for cultivation.

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.

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