Polyclonal antibody, 7696 antiserum raised against human recombinant calretinin (Swart, Bellinzona, Switzerland) and Chemicon (AB149).
Fixation/Preparation
These antibodies are applicable to formalin-fixed paraffin sections. Pretreatment of tissue sections with citrate buffer in a microwave oven or with 0.01% pronase in phosphate-buffered saline at room temperature increases calretinin immunoreactivity (Doglioni et al, 1996).
Background
7696 antiserum was raised against human recombinant calretinin (Schwaller et al, 1993) whilst AB149 antiserum was raised against guinea pig calretinin. Calretinin is a calcium-binding protein of 29 kD. It is a member of the large family of EF-hand proteins, to which the S-100 protein also belongs. EF-hand proteins are characterized by a peculiar amino acid sequence that folds up into a helix-loop-helix that acts as a calcium-binding site. Calretinin contains six such EF-hand stretches (Rogers et al, 1990). The calretinin gene was initially isolated from a cDNA clone from the chick retina and shows 60% homology with the calbindin gene (Parmentier, 1990). It is abundantly expressed in central and peripheral neural tissues, particularly in the retina and in neurons of the sensory pathways (Schwaller et al, 1993; Andressen et al, 1993). In addition, consistent calretinin immunoreactivity is found in the normal mesothelial cell lining of all serosal membranes, eccrine glands of skin, convoluted tubules of kidney, Leydig and Sertoli cells of the testis, epithelium of rete testis, endometrium and ovarian stromal cells and adrenal cortical cells.
Applications
The polyclonal antisera to calretinin consistently immunostained mesothelial cells and malignant mesotheliomas both in routinely fixed, embedded tissue sections and in cytological preparations of serous effusions (Doglioni et al, 1996). The diagnostic sensitivity of this immunocytochemical approach reached 100%, allowing immunostaining of all 44 mesotheliomas investigated, which included five biphasic and three sarcomatoid types. The specificity of calretinin immunoreactivity was checked against 294 adenocarcinomas of different origin (19 serosal metastases and 275 primary tumors potentially capable of metastases to the serosal membranes). Only 28 cases showed focal immunoreactivity for calretinin.
Comments
It is recommended that calretinin, as a mesothelioma-binding antibody, be used in conjunction with the standard panel of antibodies used in the evaluation of malignant mesothelioma (Appendix 1.16). The further uses and limitations of calretinin await further evaluation.
References
•Andressen C, Blumcke I, Celio MR 1993. Calcium-binding proteins: selective markers of nerve cells. Cell Tissue Research 271: 181-208.
•Doglioni C, Dei Tos AP, Laurino L et al 1996. Calretinin: A novel immunocytochemical marker for mesothelioma. American Journal of Surgical Pathology 20: 1037-1046.
•Parmentier M 1990. Structure of the human cDNAs and genes coding for calbindin D28K and calretinin. Advances in Experimental Medicine and Biology 269:27-34. calretinin. Advances in Experimental Medicine and Biology 269:27-34.
•Rogers J, Khan M, Ellis J 1990. Calretinin and other CaBPs in the nervous system. Advances in Experimental Medicine and Biology 269: 195-203.
•Schwaller B, Buchwald P, Blumcke I, Celio MR, Hunziker W 1993. Characterization of a polyclonal antiserum against purified human recombinant calcium binding protein calretinin. Cell Calcium 14:639-648.
Bibliografía
Manual of diagnostic antibodies for immunohistology / Anthony S.-Y. Leong, Kumarasen Cooper, F. Joel W.-M. Leong.
DESCARGAR INFORMACION TRADUCIDA AL IDIOMA ESPAÑOL
Fixation/Preparation
These antibodies are applicable to formalin-fixed paraffin sections. Pretreatment of tissue sections with citrate buffer in a microwave oven or with 0.01% pronase in phosphate-buffered saline at room temperature increases calretinin immunoreactivity (Doglioni et al, 1996).
Background
7696 antiserum was raised against human recombinant calretinin (Schwaller et al, 1993) whilst AB149 antiserum was raised against guinea pig calretinin. Calretinin is a calcium-binding protein of 29 kD. It is a member of the large family of EF-hand proteins, to which the S-100 protein also belongs. EF-hand proteins are characterized by a peculiar amino acid sequence that folds up into a helix-loop-helix that acts as a calcium-binding site. Calretinin contains six such EF-hand stretches (Rogers et al, 1990). The calretinin gene was initially isolated from a cDNA clone from the chick retina and shows 60% homology with the calbindin gene (Parmentier, 1990). It is abundantly expressed in central and peripheral neural tissues, particularly in the retina and in neurons of the sensory pathways (Schwaller et al, 1993; Andressen et al, 1993). In addition, consistent calretinin immunoreactivity is found in the normal mesothelial cell lining of all serosal membranes, eccrine glands of skin, convoluted tubules of kidney, Leydig and Sertoli cells of the testis, epithelium of rete testis, endometrium and ovarian stromal cells and adrenal cortical cells.
Applications
The polyclonal antisera to calretinin consistently immunostained mesothelial cells and malignant mesotheliomas both in routinely fixed, embedded tissue sections and in cytological preparations of serous effusions (Doglioni et al, 1996). The diagnostic sensitivity of this immunocytochemical approach reached 100%, allowing immunostaining of all 44 mesotheliomas investigated, which included five biphasic and three sarcomatoid types. The specificity of calretinin immunoreactivity was checked against 294 adenocarcinomas of different origin (19 serosal metastases and 275 primary tumors potentially capable of metastases to the serosal membranes). Only 28 cases showed focal immunoreactivity for calretinin.
Comments
It is recommended that calretinin, as a mesothelioma-binding antibody, be used in conjunction with the standard panel of antibodies used in the evaluation of malignant mesothelioma (Appendix 1.16). The further uses and limitations of calretinin await further evaluation.
References
•Andressen C, Blumcke I, Celio MR 1993. Calcium-binding proteins: selective markers of nerve cells. Cell Tissue Research 271: 181-208.
•Doglioni C, Dei Tos AP, Laurino L et al 1996. Calretinin: A novel immunocytochemical marker for mesothelioma. American Journal of Surgical Pathology 20: 1037-1046.
•Parmentier M 1990. Structure of the human cDNAs and genes coding for calbindin D28K and calretinin. Advances in Experimental Medicine and Biology 269:27-34. calretinin. Advances in Experimental Medicine and Biology 269:27-34.
•Rogers J, Khan M, Ellis J 1990. Calretinin and other CaBPs in the nervous system. Advances in Experimental Medicine and Biology 269: 195-203.
•Schwaller B, Buchwald P, Blumcke I, Celio MR, Hunziker W 1993. Characterization of a polyclonal antiserum against purified human recombinant calcium binding protein calretinin. Cell Calcium 14:639-648.
Bibliografía
Manual of diagnostic antibodies for immunohistology / Anthony S.-Y. Leong, Kumarasen Cooper, F. Joel W.-M. Leong.
DESCARGAR INFORMACION TRADUCIDA AL IDIOMA ESPAÑOL
