ab78451 reacts specifically with CD146.
In blood and marrow ab78451 reacts only with endothelial cells, and has been used to detect circulating endothelial cells in human, dogs, rabbits and mouse samples.
ab78451 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, EBV-transformed B cell lines, 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 ab78451 are also 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. We have data to indicate that this antibody may not cross react with Rat. However, this has not been conclusively tested and expression levels may vary in certain cell lines/tissues.
Human, mouse or dog blood or marrow cells
Human intestinal smooth muscle cells
Melanoma A2058, SKMEL.3 or A375.S2 cells
Normal, primary HUVEC or MVEC cultures
Human skin, intestine, ovary tonsil, lymph node, lung or kidney tissue.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 - 10 µg/ml. Works well in frozen tissue; fixed or unfixed.
Use a concentration of 1 - 10 µg/ml. Works best on EDTA or Trypsin lifted endothelial cells.
Use 2µg for 106 cells.
Methanol fixed cells.
ab91356 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
Plays a role in cell adhesion, and in cohesion of the endothelial monolayer at intercellular junctions in vascular tissue. Its expression may allow melanoma cells to interact with cellular elements of the vascular system, thereby enhancing hematogeneous tumor spread. Could be an adhesion molecule active in neural crest cells during embryonic development. Acts as surface receptor that triggers tyrosine phosphorylation of FYN and PTK2, and a transient increase in the intracellular calcium concentration.
Detected in endothelial cells in vascular tissue throughout the body. May appear at the surface of neural crest cells during their embryonic migration. Appears to be limited to vascular smooth muscle in normal adult tissues. Associated with tumor progression and the development of metastasis in human malignant melanoma. Expressed most strongly on metastatic lesions and advanced primary tumors and is only rarely detected in benign melanocytic nevi and thin primary melanomas with a low probability of metastasis.
Overlay histogram showing MCF-7 cells stained with ab78451 (red line). The cells were fixed with methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab78451, 2µg/1x106 cells) for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 FITC used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a significantly decreased signal in MCF-7 cells fixed with 4% paraformaldehyde (10 min) used under the same conditions.
Please note that Abcam does not have data for use of this antibody on non-fixed cells. We welcome any customer feedback.