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In blood and marrow ab77928 reacts only with endothelial cells, and has been used to detect circulating endothelial cells in human, dogs, rabbits 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. ab77928 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 ab77928 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.
Our Abpromise guarantee covers the use of ab77928 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-Fr||Use a concentration of 1 - 10 µg/ml.|
|IP||Use a concentration of 1 - 10 µg/ml.|
|ELISA||Use a concentration of 1 - 10 µg/ml.|
|ICC||Use a concentration of 1 - 10 µg/ml.
Works best on EDTA or Trypsin lifted endothelial cells.
|Flow Cyt||Use a concentration of 1 - 10 µg/ml. ab18434-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.|
|IHC (PFA fixed)||Use a concentration of 1 - 10 µg/ml.
4% PFA for 30min RT or <2hrs at 4°C. Block with 1% BSA/0.2% tween20/PBC for 30 min.
ab77928 has not yet been referenced specifically in any publications.