CD146, a member of the immuoglobulin superfamily, is an intrinsic membrane glycoprotein of 110-120kD found on the surface of endothelial cells, bone marrow fibroblasts and various melanomas. CD146 has been used as a marker of tumor progression in human melanoma because expression in those tumors correlates strongly with poor prognosis and the development of metastic disease. In addition, a number of human T, B and myeloid leukemic cell lines seem to express CD146. The close structural relationship with N-CAM and related molecules suggests that CD146 may be also a developmentally regulated cell adhesion molecule (Melanoma adhesion molecule or MCAM).
In blood and marrow clone 2Q401reacts only with endothelial cells, and has been used to detect circulating endothelial cells in human, dogs and mouse samples. It positively stains normal, primary HUVEC and MVEC cultures and the endothelial cells of all vessels in normal frozen sections of human skin, intestine, ovary tonsil, lymph node, lung, and kidney. It does not stain carcinoma cell lines HT-29 and COLO205, and M21, the T cell lines Jurkat and HuT78, fibroblasts, HL-60 or CHO cells, or EBV-transformed B cell lines, although it expected that CD146 expression is present in many tumor lines. 2Q401 does not stain normal monocytes, granulocytes, red cells, platelets, T cells or B cells from marrow or peripheral blood; nor does it detect marrow megakaryocytes or the megakaryoblast line HU3. The peripheral blood cells that do stain with 2Q401 also are positive for both von Willebrand Factor (vWF) and thrombomodulin (the combined expression of which is limited to endothelium), and they stain for flt and flk. In recent testing, melanoma A2058, SKMEL.3 and A375.S2 are positive, while murine melanoma M3 is negative. One smooth muscle line (HISM) was positive and one negative (TIGHA-VSMC). Human blood lymphoctes are negative with or without prior stimulation.
Applications
Suitable for use in Flow Cytometry, ELISA, Western Blot, Immunoprecipitation, Immunohistochemistry and Immunocytochemistry. Other applications not tested.
Recommended Dilutions
Flow Cytometry: 1-10ug/ml ELISA: 1-10ug/ml Western Blot: 1:500 Immunoprecipitation: 1-10ug/ml Immunohistochemistry: 1-10ug/ml Immunocytochemistry: 1-10ug/ml Optimal dilutions to be determined by the researcher.
Storage and Stability
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Form
Supplied as a liquid in PBS, pH 7.6, 0.09% sodium azide.
Purity
Purified by Protein A affinity chromatography.
Specificity
Recognizes human CD146, Mr 110kD. In blood and marrow, reacts only with endothelial cells. Species Crossreactivity: canine, rabbit, mouse (Detects circulating endothelial cells). Does not react with rat.
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