DOUBLE STAINS

Double Stains

BCL-1(cyclin D1)/CD20

Applications:  This combination can aid in the differentiation of B-cell lymphomas from mantle cell lymphoma (cyclin D1 positive).

BCL-1 (nuclear/brown) is useful in differentiating mantle cell lymphoma from chronic lymphocytic leukemia (CLL). Both mantle cell and CLL are CD5 co-expressing B-cell lymphoproliferative disorders.

CD20 (cytoplasmic/red) is restricted to expression in normal and neoplastic B cells, and is absent in other lymphocytes and tissues.

BCL-2/BCL-6

Applications:  This combination can aid in the differentiation of B-cell non-Hodgkin lymphoma of follicular center cell origin (follicular, diffuse large B-cell and Burkitt lymphoma) from other B-cell lymphoproliferative disorders.

BCL-2 (cytoplasmic/red) is expressed in most cases of follicular lymphoma and is negative in reactive follicular hyperplasia. It is also expressed in most B-cell lymphomas and a mixture of benign and neoplastic T cells.

BCL-6 protein (nuclear/brown) is expressed primarily in cell derived from the  follicle center.  BCL-6 is not expressed in hairy cell leukemia or in mantle cell and marginal-zone derived lymphomas.

CD3/CD20

Applications:  This combination can be used to aid in distinguishing between T- and B-cell proliferations in both lymphoid and non-lymphoid tissues.

CD3 (cytoplasmic/brown) is a pan T-cell antibody that recognizes T cells in thymus, bone marrow, peripheral lymphoid tissue, and blood.  The CD3 antibody detects both normal and neoplastic T cells. It is also expressed in the cytoplasm of NK cells.

CD20 (cytoplasmic/red) is restricted to expression in normal and neoplastic B cells, and is absent in other lymphocytes and tissues.

CDX2/CK7

Applications:  This combination can be used to distinguish cancers derived from the gastrointestinal tract from those of breast, lung, and ovary.

CDX2 (nuclear/brown) is an intestine-specific transcription factor.  It is expressed in the nuclei of epithelial cells throughout the intestine, from duodenum to rectum.  The CDX2 protein is expressed in primary and metastatic colorectal carcinomas.
 
CK7 (cytoplasmic/red) is a basic cytokeratin and is expressed in epithelial cells and cacners of the ovary, lung, and breast, but not in the colon or lower gastrointestinal tract.

Cytokeratin HMW/LMW

Applications:  This combination can be used to differentiate between squamous cell carcinomas and adenocarcinomas.

CK-HMW (34bE12 clone) (cytoplasmic/brown) reacts with cytokeratin (CK) 1, 5, 10, and 14.  It is reactive with stratified epithelia, myoepithelial, and basal cells of the prostate and breast.  It stains squamous cell carcinomas and adenosquamous carcinomas.

CK-LMW (clone 5D3) (cytoplasmic/red) recognizes cytokeratins 8 (CK8) and 18 (CK18) of simple and glandular epithelium and adenocarcinomas.

D2-40 / BerEp4

Applications: This combination is designed to specifically consider the diagnosis of mesothelioma versus adenocarcinoma. This combination of antigens was selected for the combination of sensitivity, specificity and robustness of staining patterns.

D2-40 (also known as podoplanin) has a broad range of reactivities including lymphatic endothelium and mesothelial cells. It is positive in the majority of mesotheliomas.

BerEp4 is expressed in adenocarcinomas. It is both sensitive and specific and is especially useful in distinguishing mesothelioma from adenocarcinoma.  
    

Melan-A/Ki67

Applications:   This double stain can be used to differentiate malignant melanoma from melanocytic nevi manifesting cellular and/or structural atypia by virtue of a significant difference in the proportion of Ki67-positive cells.  Malignant melanomas have higher proliferative activity than melanocytic nevi in the deeper dermis, although this finding should be interpreted with caution.

Melan-A (cytoplasmic/red) is expressed in 100% of melanocytes, most melanomas, and 50% to 60% of melanoma cell lines.

Ki67 is a nuclear protein (nuclear/brown) expressed in proliferating cells.  Ki67 is preferentially expressed during late G1-, S-, M-, and G2-phases of the cell cycle, while cells in the G0 (quiescent) phase are negative for this protein.

 

PAX 5 / CD5

Applications:  This combination aids in the differentiation of CD5 co-expressing B-cell lymphoproliferative disorders from other mature B- or T-cell processes.

PAX 5 (nuclear/brown) is expressed primarily in pro-, pre-, and mature B cells, but not in plasma cells. In contrast to CD20, PAX5 expression is not down-regulated after therapy with rituximab.

CD5 (cytoplasmic/red), a T-cell marker expressed in many T-cell leukemias and lymphomas. It is also expressed in mantle cell lymphoma, chronic lymphocytic leukemia/small lymphocytic lymphoma and a very small subset of normal B cells.
    

PAX 5 / CD43

Applications:  This combination can aid in the differentiation of B-cell lymphoproliferative disorders from other mature B- or T-cell processes.

PAX 5 (nuclear/brown) is expressed primarily in pro-, pre-, and mature B cells, but not in plasma cells.

CD43 (cytoplasmic/red) identifies T cells and cells of myeloid lineage.  It is useful in the diagnosis of T-cell lymphoma.  Aberrant expression of CD43 labels a subset of B-cell lymphoproliferative disorders.