Immunohistochemistry Antibody Specificity | College of Veterinary Medicine

Immunohistochemistry Antibody Specificity

Intermediate Filaments

Cytokeratin (AE1/AE3)

This particular antibody labels high and low molecular weight keratins in many epithelia, including keratinized and corneal epidermis, stratified squamous epithelia of internal organs, stratified epithelia, hyperproliferative keratinocytes, and simple epithelia.

Vimentin

Vimentin is present in cells of mesenchymal origin and in mesenchymal derived tumors such as lymphomas, sarcomas and melanomas.

Desmin

Antibodies against desmin label smooth and striated muscle cells and is useful in identification of tumors of myogenic origin such as leiomyosarcomas and rhabdomyosarcomas.

Smooth Muscle Actin (SMA)

The smooth muscle actin protein is present in smooth muscle and immunoreactivity for SMA is seen in leiomyosarcomas and leiomyomas.

Muscle Specific Actin (HHF-35)

This antibody recognizes muscle specific alpha and gamma actin isomers but is not reactive to beta isomers. It will stain myocardial, smooth and skeletal muscle cells as well as myoepithelial cells and pericytes of small vessels.

Keratin 8

Cytokeratin 8 is primarily found in non squamous epithelium and is present in the majority of adenocarcinomas and ductal carcinomas.

Adhesion

E-Cadherin

E-cadherin plays a crucial role in the adherens junction in epithelium and is important in maintaining epithelial integrity. Down-regulation of E-cadherin expression has been observed in carcinomas. Expression of E-cadherin may be important in determining the invasive potential of epithelial neoplasms and the transition from benign to malignant neoplasms.

Desmoplakin

Desmoplakin is a key component of the desmosome and anchors the intermediate filament cytoskeleton to the cell membrane. Reduced expression of desmoplakin has been correlated with the progression of several cancers.

Hematopoietic

CD3

The CD3 antigen is present on the cell membrane of normal T cells and is expressed in the majority of T-cell neoplasms.

CD20

CD20 is expressed in normal and neoplastic B cells.

CD79a

This antigen is expressed in all B-cell neoplasms. Artefactual nuclear staining is common with this antibody.

CD18 (dog)

CD18 is expressed by most leukocytes. Defects in the CD18 gene results in leukocyte adhesion deficiency. Although highly expressed in neoplasms of histiocytic origin, CD18 may also be identified in other hematopoietic neoplasms. This antibody only works in the canine.

CD31

Antibody against CD31 strongly labels endothelial cells and neoplasms derived from endothelial cells.

CD34

The antibody against CD34 reacts with hematopoietic precursor cells and endothelial cells. Acute myeloid leukemias, acute lymphoblastic leukemias, undifferentiated leukemias, and CML blasts are positive with this antibody. This is a good marker for endothelial cells in the mouse and can be used on formalin-fixed, paraffin-embedded material.

CD45

CD45 is present on the cell membrane of normal and neoplastic leukocytes. CD 45 is also referred to as Leucocyte Common Antigen. This antibody reacts specifically with human CD45 and is useful for identifying human leukocytes transplanted into immunodeficient mice.

CD163

This antibody reacts with most of macrophages in lymphoreticular organs and in many other organs and tissues. This antibody does not react with dendritic cell population, such as epidermal Langerhans cells, interdigitating cells in the paracortex of lymph nodes, nor follicular dendritic cells. This antibody recognizes the antigen presenting on the cell surface membrane of tissue macrophages.

Lambda light chain

This antibody stains only lambda light chain-containing cells in lymphoid tissue. Since most plasma cell tumors in the dog express lambda rather than kappa light chains, it is a good marker for plasma cell tumors in the dog. Elevated levels of free light chain occur in multiple myelomas and other B-cell malignancies.

Neural / Melanoma

Melan-A

MelanA is expressed in the majority of melanomas as well as in melanocytes. Also referred to as MART1 (Melanoma Antigen Recognized by T cells).

S-100

The S100 antibody may be useful for the identification of malignant neoplasms, including chondroblastomas, schwannomas, and melanomas; however, many cell types are S-100-positive.

Synaptophysin

Synaptophysin is expressed by normal neuroendocrine cells and neuroendocrine neoplasms.

Chromogranin

Chromogranin is expressed by normal cells and neoplasms of neuroendocrine origin.

Neuron Specific Enolase

Neuron-specific enolase is expressed by cells of neuronal and neuroendocrine origin.

GFAP

Antibodies against Glial Fibrillary Acidic Protein label astrocytes and some CNS ependymal cells.

Miscellaneous

Factor VIII-related antigen

Factor VIII-realted antigen (vonWillebrand factor) is present in endothelial cells and in the cytoplasm of megakaryocytes. This antibody identifies normal and neoplastic vascular endothelial cells and megakaryocytic proliferation.

Herpes Simplex-1

This antibody reacts with type specific as well as type-common antigens. It can be used to identify HXV-1 in the tissues of many species.

PCNA

PCNA is a marker for cells in early G1 and S phase of the cell cycle.

c-Kit

Antibodies against c-Kit label breast epithelium, germ cells, melanocytes, stem cells and mast cells. It has been shown to label testicular germ cell neoplasms, endometrial carcinomas, papillary and follicular thyroid carcinomas, small cell carcinomas, melanomas and ovarian epithelial carcinomas. Reactivity for c-kit may be of prognostic value in mast cell tumors of the dog.

Cleaved Caspase 3

The caspase family of cysteine proteases plays an important role in apoptosis. Cytoplasmic immunoreactivity for cleaved caspase 3 is considered a marker for apoptosis.

Mac387

Mac387 is expressed by granulocyte, monocytes and tissue macrophages and tumors derived from these cells in a variety of species.

Mouse Specific

B-220 (CD45R)

This antibody in B lymphocytes at all stages of maturation, but is decreased on plasma cells and a subset of memory B cells. It is also found on abnormal T cells and on lytically active subsets on NK cells. This antibody is mouse specific but cross reaction with human T lymphocytes has been observed.

Keratin 6

Keratin 6 is found in hair follicles and squamous cell carcinomas. In the epidermis, it is considered a marker for prolifearion.

Keratin 10

Cytokeratin 10 is present in suprabasal layers of keratinizing stratified epithelium and in differentiated areas of squamous cell carcinoma.

Keratin 14

Cytokeratin 14 is present in the basal layer of stratifying squamous and non-squamous epithelia. Its expression is increased in squamous cell carcinoma.

CD45R (frozen)

The antibody against CD45 reacts with B cells, T cell subsets, monocytes, macrophages and weakly with granulocytes.

CD4 (frozen)

CD4 is highly expressed on a subset of T lymphocytes and, at low levels, on monocytes, tissue macrophages and granulocytes.

CD8 (frozen)

The CD8 cell surface glycoprotein is found on most cytotoxic T lymphocytes.

F4/80 (Macrophage)

Murine macrophages express F4/80 antigen.

Insulin

This antibody is is well-suited for the detection of insulin in a variety of species.

Lysozyme

This antibody is useful for the identification of histiocytic neoplasias and myeloid leukemias.

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Ly6G (neutrophil)

Ly6G is expressed predominantly on neutrophils.

Comparative Pathology and Mouse Phenotyping Shared Resource