New antiserum against Ki-67 antigen suitable for double immunostaining of paraffin wax sections.
OBJECTIVE
To prepare a rabbit antiserum equivalent to MIB 1 to permit the simultaneous assessment of cell proliferation and other markers of interest using double labelling studies.


METHODS
Rabbits were immunised with a synthetic peptide deduced from the cDNA sequence coding for the Ki-67 antigen. Serum samples were tested for immunoreactivity using different immunobiochemical methods.


RESULTS
A polyclonal antiserum was derived which detects the native as well as recombinant parts of the Ki-67 antigen in different test systems. Furthermore, the antiserum stains the Ki-67 antigen in routinely processed, paraffin wax embedded material.


CONCLUSIONS
After antigen unmasking by microwave treatment the antiserum described here represents a powerful tool for the determination of growth fractions even in archival material. It is especially suitable for double staining experiments in combination with monoclonal antibodies.
Key
G
G
Forschungsinstitut Borstel, Division of Molecular Immunology, Germany.
Petersen
J L
JL
Becker
M H
MH
Duchrow
M
M
Schlüter
C
C
Askaa
J
J
Gerdes
J
J
D000595:Amino Acid Sequence; D000818:Animals; D002455:Cell Division / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry / Q000276:immunology*; D007106:Immune Sera / Q000276:immunology*; D007150:Immunohistochemistry; D007202:Indicators and Reagents; D019394:Ki-67 Antigen; D008214:Lymphocytes / Q000276:immunology; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000737:chemistry / Q000276:immunology*; D009687:Nuclear Proteins / Q000737:chemistry / Q000276:immunology*; D016612:Paraffin Embedding; D011817:Rabbits
D000939:Epitopes; D007106:Immune Sera; D007202:Indicators and Reagents; D019394:Ki-67 Antigen; D009363:Neoplasm Proteins; D009687:Nuclear Proteins