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PRINCIPLE OF THE TEST.
Used in immunohistochemistry for proteolytic digestion.
SPECIMEN REQUIRED.
All human tissue excised at surgery, outpatient clinics, and postmortems,
fresh or in fixative, along with a filled-out Tissue Examination Form
(U. S. Standard Form 515, USSF515).
REAGENTS, INSTRUMENTATION.
Trypsin.
Calcium chloride.
Deionized water.
Sodium hydroxide.
STEP-BY-STEP DESCRIPTION.
SOLUTION:
A. Trypsin........................ 50mg
B. Calcium chloride............... 50mg
C. Deionized water................ 50ml
Adjust pH to 7.8 with sodium hydroxide.
PROCEDURE:
A. Cut section using standard procedure for immunopath.
B. Allow slides to dry and then deparaffinize and bring to water.
C. Incubate sections in the trypsin
in a 37o C oven for 30 minutes.
D. Rinse sections for 5 minutes with running tap water.
E. Rinse sections twice with PBS.
F. Continue with the routine peroxidase procedure.
REFERENCES.
1. Miller KD.
Immunohistochemical Techniques. Chapter 21. pp. 421-464.
In: Bancroft JD, Gamble M.
Theory and Practice of Histological Techniques. Fifth Edition.
Edinburgh: Churchill Livingstone. 2002;:421-464.
ISBN 0-443-06435-0, 796 pages.
2. Prophet EB, Mills B, Arrington JB, Sobin LH, eds.
Laboratory Methods in Histotechnology.
Washington, DC: Armed Forces Institute of Pathology. 1992;:.
ISBN 1-881041-00-X, 278 pages.
3. Mikel UV, ed.
Advanced Laboratory Methods in Histology and Pathology.
Washington, DC: Armed Forces Institute of Pathology. 1994;:.
ISBN 1-881041-13-1, 254 pages.