sigma弗氏不*佐剂

FREUND'S ADJUVANT, COMPLETE AND

INCOMPLETE

Product Number F 5881 AND F 5506

Storage Temperature 2-8 °C

Product Description

Appearance

F 5881 Clear amber liquid containing particulate matter

（dried cells）

F 5506 Clear amber liquid

Freund's Adjuvant is one of the most commonly used

adjuvants in research. It is used as a water-in-oil

emulsion. It is prepared from non-metabolizable oils

（paraffin oil and mannide monooleate）. If it also

contains killed Mycobacterium tuberculosis it is known

as Complete Freund's Adjuvant. Without the bacteria it

is Incomplete Freund's Adjuvant. First developed by

Jules Freund in the 1940's, Freund's Adjuvant is

designed to provide continuous release of antigens

necessary for stimulating a strong, persistent immune

response1,2,3 The main disadvantage of Freund's

Adjuvant is that it can cause granulomas, inflammation

at the inoculation site and lesions. The mycobacteria in

Complete Freund's attracts macrophages and other

cells to the injection site which enhances the immune

response. For this reason, the Complete Freund's

Adjuvant is used for the initial injections. To minimize

side-effects, Incomplete Freund's Adjuvant is used for

the boosts.

For comparisons of different adjuvant systems, see

references 4 and 5.

Reagents

Each ml of F 5881 contains 1 mg of heat-killed and

dried Mycobacterium tuberculosis （strain H37Ra, ATTC

25177）, 0.85 ml paraffin oil and 0.15 ml of mannide

monooleate.

Each ml of F 5506 contains 0.85 ml of paraffin oil and

0.15 ml of mannide monooleate.

Precautions and Disclaimer

Please consult the Material Safety Data Sheet for

handling recommendations before working with this

material.

Storage/Stability

Store in a cooler at 2-8 °C. Do not Freeze.

Procedure

1. If using Complete Freund’s Adjuvant, vortex or

shake to resuspend the Mycobacterium.

2. Mix antigens （preferably in saline） with an equal

volume of the adjuvant to form an emulsion. In

order to do this, vigorous and prolonged mixing is

needed. There are at least three methods which

can be used to accomplish this:

For small volumes the emulsion can be made in a

tube. Pipet the adjuvant in the tube first. Then,

while vortexing, add an equal volume of the antigen

solution. Vortex vigorously until a thick emulsion

forms.

For intermediate volumes, use two syringes

connected through a luer fitting. Ideally, a 3-way

valve should be used. Take the desired amount of

antigen solution into a glass syringe. The volume

should not fill more than half the syringe. Take an

equal volume of the adjuvant into another glass

syringe. Remove all air and connect the syringes

through the luer fitting to the 3-way valve. Adjust

the 3-way valve such that the connection is open

between the two syringes. Carefully depress the

plunger from the antigen solution first, pushing the

antigen into the oil of the adjuvant. Alternay push

the plungers, mixing the adjuvant and the antigen

solution into an emulsion. Continue until the

plungers are difficult to push.

For large volumes, use a tissue homogenizer to

make the emulsion. Add the adjuvant to the

homogenizer first. Run the homogenizer for a short

time to coat the inside with the adjuvant. Add an

equal volume of the antigen solution and run until a

thick emulsion forms.

3. The resulting emulsion should be very thick and a

drop of it should not disperse if tested by placing on

the surface of a saline solution.

4. Transfer the emulsion to a syringe （or remove one

syringe from the luer fitting if using the two-syringe

method）. Remove all the air. Add an appropriay

sized needle. The samples are now ready for

injection.6

sigma弗氏不*佐剂

References

1. Freund, J. and McDermott, K., Proc. Soc. Exp. Biol.

Med., 49, 548-553 （1942）

2. Freund, J., Ann. Rev. Microbiol., 1, 291 （1947）

3. Freund, J., Adv. Tuberc. Res., 7, 130 （1956）

4. Bennett, B. et al., J. Immuno. Meth., 153, 31-40

（1992）

5. Deeb, B.J. et al., J. Immuno. Meth., 152, 105-113

（1992）

6. Harlow, E. and Lane, D., Antibodies A Laboratory

Manual, （Cold Spring Harbor Laboratory, 1988）

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