Immunocytohistochemistry Flashcards
Produced by immunizing an animal with a purified specific molecule (immunogen) that contains the antigen of interest,
collecting Ig-rich serum after the animal produces humoral antibodies.
Polyclonal antibodies
Products of an individual clone of plasma cells
Pruduced through hybridoma and cloning techniques
Monoclonal antibodies
absolute methanol, or acetone
to preserve immunological activity
prevent destruction of some labile antigenic site
Cryostat section
immuno flluorescence
immunoperoxidase techniques
Formaldehyde-fixed and paraffin embedded tissues
highly sensitive, present in epithelial tumors (carcinoma)
Keratin
carcinomas of lung, breast, uterus, and ovaries (serous tumors)
CK7 (Cytokeratin 7
carcinomas of colon and stomach
CK20
Transitional Cell carcinomas of the bladder and mucinous ovarian tumors
CK7 and CK20 –
high molecular weight protein that is helpful in determining the site of tumor.
It is positive for adenocarcinomas of the breast, lungs, and kidneys
Epithelial Membrane Antigen (EMA)
oncofetal antigen present in carcinomas of the GIT, pancreas, lung, breast, ovary, uterus, and cervix.
It is useful for differentiating adenocarcinoma (+) and mesothelioma (-)
Carcinoembryonic Antigen (CEA)
useful in distinguishing lung adenocarcinomas from mesotheliomas
Thyroid Transcription Factor-1 (TTF-1)
useful in the diagnosis of prostatic adenocarcinoma
Prostate-Specific Antigen (PSA)
sensitive marker for muscle differentiation and can be used to identify tumors derived from muscles
Actin
a 57kD intermediate filament that is present in normal mesenchymal cells and their neoplastic counterparts
melanomas and schwannomas stain(+)
Vimentin
a 53kD intermediate filament expressed by smooth and striated muscles. It is highly specific for myogenic tumors.
Desmin
a 51kD intermediate filament protein expressed by CNS glial cells, particularly astrocytes
Glial fibrillary acidic protein (GFAP)
expressed in cells of neural origin, particularly neurons, neuronal processes, peripheral nerves, sympathetic ganglia, adrenal medulla, and neuroendocrine cells
Neurofilament (NF)
low-molecular weight calcium-binding protein that is expressed in CNS glial cells, Schwann cells, melanocytes, histiocytes, chondrocytes, skeletal and cardiac muscle, myoepithelial cells, and some epithelial cells of breast, salivary, and sweat glands
S100 Protein
provides strong evidence of neural or neuroendocrine differentiation
2.
Neuron-specific enolase (NSE)
a marker for neuroendocrine differentiation
Chromogranin
a 38kD transmembrane protein associated with presynaptic vesicles of neurons
Synaptophysin
synthesized by placental syncytiotrophoblasts
a marker for choriocarcinoma
Human Chorionic Gonadotropin (HCG)
marker for endodermal sinus tumors showing yolk sac differentiation
Alpha-fetoprotein (AFP)
marker for germ cell tumors, particularly germinomas
Placenta-like alkaline phosphatase (PLAP)
positive for muscle-specific actin, desmin, myo D1, myoglobin, and myogenin
Myogenic Tumors
CD 68, FAM 56, alpha-1-antitrypsin, alpha-1-antichymotrypsin
Fibrohistiocytic tumors
Factor VII-related antigen, CD 31, Ulex europaeus 1 (UEA)
Vascular Tumors
S100 protein, melanosome (HMB-45), Melan-A (MART-1)
Melanomas
– Leukocyte Common Antigen (LCA) or also called CD 45.
Lymphomas
- T cell : CD3, CD4, CD8
- B cells: CD19, CD20, CD23
- Reed-Sternberg cells: CD15, CD30,
- Ig light and heavy chains
Immunophenotypic, subclass
enzyme label
INSOLUBLE DARK BROWN REACTION
Optimal incubation time =
Horseradish peroxidase + Diaminobenzidine
30-60 minutes @ room temp.
most common immunoglobulin
IgG
most common animal for monoclonal Ab prod
mice
most common animal for polyclonal Ab prod
rabbit
Most common enzyme label
peroxidase
DAM (diaminobenzidine)
brown
AEC (aminoethylcarbazole)
red
