Sources/Clones
Triton Diagnostics and Zymed (MAK-6 - clones KA4 and UCD/PR10.11).
Fixation/Preparation
MAK-6 works well in routinely fixed, paraffin-embedded tissue sections. Trypsin pretreatment is necessary for antigen unmasking. Incubation of the primary antibody for 1 h at 37 C or overnight incubation at room temperature yields superior immunostaining. Preincubated with blocking reagents to reduce non-specific background staining has been recommended, but we have found this to be unnecessary.
Background
MAK-6 antibody cocktail contains an optimized mixture of two murine monoclonal antibodies of IgG1 isotype. Antibody kA 4 recognizes human cytokeratin types, 14, 15, 16 and 19 while antibody UCD/PR-10.11 recognizes human cytokeratins 8 and 18.
Antibody UCD/PR 10.11 was produced using shed extracellular antigen purified from MCF-7 tissue culture media and was selected for its specificity to cytokeratin types 8 and 18 (Chan et al, 1986). Antibody kA4 was produced against human sole epidermis and was selected for its specificity to cytokeratin types 14, 15, 16 and 19.
Applications
MAK-6 is reputed to stain all cases of squamous cell carcinomas and the majority of adenocarcinomas, carcinoid tumors and undifferentiated carcinomas. Lymphomas, melanomas, gliomas/astrocytomas and the majority of sarcomas do not demonstrate MAK-6 positivity. The latter is related to the expected cytokeratin expression in synovial sarcomas and epithelioid sarcomas. It should be noted that the majority of ependymomas and basal cell carcinomas of the skin also do not express MAK-6 Caution should be observed in assessing metastatic carcinomas to the brain, since MAK-6 may rarely show crossreactivity with neural tissue. In these instances application of antibody to glial fibrillary acidic protein (GRAP) would be helpful (Cooper et al, 1985; McNutt et al, 1988).
Comments
We have found MAK-6 to be a useful pankeratin marker. Strong immunoreaction is demonstrated in tissue of epithelial origin. Used in conjunction with other pankeratin markers, the majority of neoplasms showing cytokeratin expression may be identified.
References
•Chan R, Rossitto PV, Edwards BF, Cardiff RD 1986. Presence of proteolytically processed keratins in the culture medium of MCF-7 cells. Cancer Research 46 (1): 6353-6359.
•Cooper D, Schermer A, Sun T-T 1985. Classification of human epithelia and their neoplasms using monoclonal antibodies to keratins: strategies, applications and limitations. Laboratory Investigation 52: 243-256.
•McNutt MA, Bolen JW, Vogel AM et al 1988 Monoclonal antibodies to cytokeratins in diagnostic immunocytochemistry. In: Wick MR, Siegel GP (eds) Monoclonal antibodies in diagnostic immunohistochemistry. New York: Marcel Dekker, pps 51-70.
Bibliografía
Manual of diagnostic antibodies for immunohistology / Anthony S.-Y. Leong, Kumarasen Cooper, F. Joel W.-M. Leong.
Triton Diagnostics and Zymed (MAK-6 - clones KA4 and UCD/PR10.11).
Fixation/Preparation
MAK-6 works well in routinely fixed, paraffin-embedded tissue sections. Trypsin pretreatment is necessary for antigen unmasking. Incubation of the primary antibody for 1 h at 37 C or overnight incubation at room temperature yields superior immunostaining. Preincubated with blocking reagents to reduce non-specific background staining has been recommended, but we have found this to be unnecessary.
Background
MAK-6 antibody cocktail contains an optimized mixture of two murine monoclonal antibodies of IgG1 isotype. Antibody kA 4 recognizes human cytokeratin types, 14, 15, 16 and 19 while antibody UCD/PR-10.11 recognizes human cytokeratins 8 and 18.
Antibody UCD/PR 10.11 was produced using shed extracellular antigen purified from MCF-7 tissue culture media and was selected for its specificity to cytokeratin types 8 and 18 (Chan et al, 1986). Antibody kA4 was produced against human sole epidermis and was selected for its specificity to cytokeratin types 14, 15, 16 and 19.
Applications
MAK-6 is reputed to stain all cases of squamous cell carcinomas and the majority of adenocarcinomas, carcinoid tumors and undifferentiated carcinomas. Lymphomas, melanomas, gliomas/astrocytomas and the majority of sarcomas do not demonstrate MAK-6 positivity. The latter is related to the expected cytokeratin expression in synovial sarcomas and epithelioid sarcomas. It should be noted that the majority of ependymomas and basal cell carcinomas of the skin also do not express MAK-6 Caution should be observed in assessing metastatic carcinomas to the brain, since MAK-6 may rarely show crossreactivity with neural tissue. In these instances application of antibody to glial fibrillary acidic protein (GRAP) would be helpful (Cooper et al, 1985; McNutt et al, 1988).
Comments
We have found MAK-6 to be a useful pankeratin marker. Strong immunoreaction is demonstrated in tissue of epithelial origin. Used in conjunction with other pankeratin markers, the majority of neoplasms showing cytokeratin expression may be identified.
References
•Chan R, Rossitto PV, Edwards BF, Cardiff RD 1986. Presence of proteolytically processed keratins in the culture medium of MCF-7 cells. Cancer Research 46 (1): 6353-6359.
•Cooper D, Schermer A, Sun T-T 1985. Classification of human epithelia and their neoplasms using monoclonal antibodies to keratins: strategies, applications and limitations. Laboratory Investigation 52: 243-256.
•McNutt MA, Bolen JW, Vogel AM et al 1988 Monoclonal antibodies to cytokeratins in diagnostic immunocytochemistry. In: Wick MR, Siegel GP (eds) Monoclonal antibodies in diagnostic immunohistochemistry. New York: Marcel Dekker, pps 51-70.
Bibliografía
Manual of diagnostic antibodies for immunohistology / Anthony S.-Y. Leong, Kumarasen Cooper, F. Joel W.-M. Leong.
