Leukemia Inhibitory Factor from mouse

RECOMMENDED QC PROTOCOL

M1 Bioassay

1. The M1 bioassay is performed using in vitro semi-solid agar cultures, which contain approximately 100 cells in 1 mL volumes of DME containing 20 % FCS in 0.3% agar.

2. Add 100 μL of sample or mLIF (10(E4) units/mL in 5% FCS in isotonic saline) in two-fold serial dilutions in duplicate to 35 mm petri dishes.

3. Add 100 μL of 5% FCS in isotonic saline to two control slides.

4. Incubate at 37°C in fully humidified atmosphere of 10% CO2 in air for 7 days.

5. Score the number of colonies that show differentiation (note: 50 units is defined as the amount of activity which results in 50% of the colonies being differentiated).

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Manufactured by CHEMICON International, Inc. LIF is protected under US Patent nos. 5,187,077, 5,427,925, 5,443,825, 5,750,654 and 6,261,548, European Patent no. 0285 448 and related foreign patents and is not available for resale.

Specific Activity: The activity of mouse LIF is determined by the ability to induce differentiation of murine M1 myeloid leukemic cells.The minimum detectable concentration of mouse LIF in this assay is 0.5 ng/mL

Tested negative in both aseptic and microplasmic tests.

Specific Activity: where 50 units is defined as the amount of mouse LIF required to induce differentiation in 50% of the M1 colonies in 1 mL agar cultures.

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