sigma弗氏不*佐剂

产品型号：F5506

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sigma弗氏不*佐剂Biochem/physiol Actions 弗氏佐剂可用于制备免疫原的油包水乳液。由于抗原释放缓慢，油包水乳液中的抗原可刺激高效长期的抗体应答。Other Notes 每毫升含 0.85mL 石蜡油和 0.15mL 二缩甘露醇一油酸。

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sigma弗氏不*佐剂

FREUND'S ADJUVANT, COMPLETE ANDINCOMPLETEProduct Number F 5881 AND F 5506Storage Temperature 2-8 °CProduct DescriptionAppearanceF 5881 Clear amber liquid containing particulate matter（dried cells）F 5506 Clear amber liquidFreund's Adjuvant is one of the most commonly usedadjuvants in research. It is used as a water-in-oilemulsion. It is prepared from non-metabolizable oils（paraffin oil and mannide monooleate）. If it alsocontains killed Mycobacterium tuberculosis it is knownas Complete Freund's Adjuvant. Without the bacteria itis Incomplete Freund's Adjuvant. First developed byJules Freund in the 1940's, Freund's Adjuvant isdesigned to provide continuous release of antigensnecessary for stimulating a strong, persistent immuneresponse1,2,3 The main disadvantage of Freund'sAdjuvant is that it can cause granulomas, inflammationat the inoculation site and lesions. The mycobacteria inComplete Freund's attracts macrophages and othercells to the injection site which enhances the immuneresponse. For this reason, the Complete Freund'sAdjuvant is used for the initial injections. To minimizeside-effects, Incomplete Freund's Adjuvant is used forthe boosts.For comparisons of different adjuvant systems, seereferences 4 and 5.ReagentsEach ml of F 5881 contains 1 mg of heat-killed anddried Mycobacterium tuberculosis （strain H37Ra, ATTC25177）, 0.85 ml paraffin oil and 0.15 ml of mannidemonooleate.Each ml of F 5506 contains 0.85 ml of paraffin oil and0.15 ml of mannide monooleate.Precautions and DisclaimerPlease consult the Material Safety Data Sheet forhandling recommendations before working with thismaterial.Storage/StabilityStore in a cooler at 2-8 °C. Do not Freeze.Procedure1. If using Complete Freund’s Adjuvant, vortex orshake to resuspend the Mycobacterium.2. Mix antigens （preferably in saline） with an equalvolume of the adjuvant to form an emulsion. Inorder to do this, vigorous and prolonged mixing isneeded. There are at least three methods whichcan be used to accomplish this:For small volumes the emulsion can be made in atube. Pipet the adjuvant in the tube first. Then,while vortexing, add an equal volume of the antigensolution. Vortex vigorously until a thick emulsionforms.For intermediate volumes, use two syringesconnected through a luer fitting. Ideally, a 3-wayvalve should be used. Take the desired amount ofantigen solution into a glass syringe. The volumeshould not fill more than half the syringe. Take anequal volume of the adjuvant into another glasssyringe. Remove all air and connect the syringesthrough the luer fitting to the 3-way valve. Adjustthe 3-way valve such that the connection is openbetween the two syringes. Carefully depress theplunger from the antigen solution first, pushing theantigen into the oil of the adjuvant. Alternay pushthe plungers, mixing the adjuvant and the antigensolution into an emulsion. Continue until theplungers are difficult to push.For large volumes, use a tissue homogenizer tomake the emulsion. Add the adjuvant to thehomogenizer first. Run the homogenizer for a shorttime to coat the inside with the adjuvant. Add anequal volume of the antigen solution and run until athick emulsion forms.3. The resulting emulsion should be very thick and adrop of it should not disperse if tested by placing onthe surface of a saline solution.4. Transfer the emulsion to a syringe （or remove onesyringe from the luer fitting if using the two-syringemethod）. Remove all the air. Add an appropriaysized needle. The samples are now ready forinjection.6sigma弗氏不*佐剂

References1. 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 LaboratoryManual, （Cold Spring Harbor Laboratory, 1988）

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