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CD146 Monoclonal Antibody (P1H12), NovaFluor™ Yellow 730, eBioscience™
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FIGURE: 1 / 2
CD146 Antibody (H058T02Y07) in Flow
Normal human peripheral blood cells were unstained (left) or stained with CD146 Monoclonal Antibody, NovaFluor Yellow 730 (Product #: h058t03y07) (right). All cells were co-stained with CD3 Monoclonal Antibody, eFluor 450 (Product #: 48-0037-42). Total viable cells in the lymphocyte gate were used for analysis, as determined by LIVE/DEAD Blue (Product # L34962). Data was acquired on a 5-laser Cytek Aurora and unmixed with autofluorescence extraction.
Product Details
H058T02Y07
Product Specifications
Host/Isotype
Mouse
/ IgG1, kappa
Class
Monoclonal
Type
Antibody
Clone
P1H12
Conjugate
Excitation/Emission Max
551/730 nm
View spectra
Form
Liquid
Concentration
0.2 µg/Test
Storage conditions
4°C, store in dark, DO NOT FREEZE!
Shipping conditions
Ambient (domestic); Wet ice (international)
RRID
Product Specific Information
Description: The monoclonal antibody P1H12 recognizes CD146 also known as MUC18, s-endo, Endo-CAM and Mel-CAM, which is a member of the Ig superfamily of proteins. The expression of CD146 is found on endothelial cells, bone marrow fibroblasts and some tumors (especially melanoma). Recently mesenchymal stromal cells and endometrial stromal cells have also been shown to express CD146. The presence of CD146 on circulating blood cells have been confined to a subset of T cells rather than circulating endothelial cells, as expression of other endothelial markers (CD31 and CD51/61) is negative. Expression can be found on activated lymphocytes. The protein is heavily glycosylated with more than 50% of the mass from carbohydrates. The antibody P1H12 has been reported to crossreact to mouse, rabbit, canine, but not rat.
Applications Reported: This P1H12 antibody has been reported for use in flow cytometric analysis.
Applications Tested: This P1H12 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells and Human Umbilical Vein Cells (HUVEC). This can be used at 5 µL (0.02 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.
Our internal testing shows that NovaFluor Yellow 730 non-specifically stains B cells in SJL mice. Non-specific staining has not been observed in BALB/c or C57BL/6 mice. Other strains have not been tested. See the Antibody Testing Data for an example of this strain-dependent difference.
NovaFluor dyes are not compatible with DNA intercalating viability dyes. Do not use viability dyes such as propidium iodide, 7-actinomycin D (7-AAD) and DAPI. Invitrogen LIVE/DEAD Fixable Dead Cell stains are recommended for use with NovaFluor dyes.
Each NovaFluor conjugate or kit is shipped with CellBlox Blocking Buffer. Use this buffer whenever staining with NovaFluor conjugates, including single-color compensation controls using cells. Whenever possible, we recommend adding CellBlox Blocking Buffer to antibody cocktails/master mixes prior to combining with cells. Add 5 µL per sample (regardless of the number of NovaFluors in your panel) to use the antibody cocktail as intended. For single-color controls, use 5 µL of CellBlox Blocking Buffer per 100µL of cell sample containing 10^3 to 10^8 cells.
NovaFluor conjugates are based on Phiton™ technology utilizing novel nucleic acid dye structures that allow for engineered fluorescent signatures with consideration for spillover and spread impacts. Learn more
Excitation: 552 nm; Emission: 730 nm; Laser: 561 nm (Yellow) Laser
Target Information
CD146, also known as MCAM (Melanoma Cell Adhesion Molecule), is an integral membrane glycoprotein belonging to the immunoglobulin superfamily. It is heavily glycosylated, with more than 50% of its mass derived from carbohydrates. CD146 is primarily expressed on endothelial cells at cell-cell junctions, playing a crucial role in cell adhesion and the cohesion of the endothelial monolayer in vascular tissue. In addition to endothelial cells, CD146 is expressed on bone marrow fibroblasts, mesenchymal stromal cells, endometrial stromal cells, and some tumors, particularly melanoma. It has also been observed on a subset of circulating T cells and activated lymphocytes, but not on circulating endothelial cells, as indicated by the absence of other endothelial markers such as CD31 and CD51/61. CD146 functions as a calcium-independent cell adhesion molecule and may also act as a signal transduction molecule. It is involved in the recruitment of Fyn kinase and the subsequent tyrosine phosphorylation of intracellular proteins, which are important for actin cytoskeleton assembly. This signaling role includes triggering tyrosine phosphorylation of FYN and PTK2 and causing a transient increase in intracellular calcium concentration. In the context of cancer, CD146 expression may facilitate the interaction of melanoma cells with the vascular system, enhancing hematogenous tumor spread. Additionally, CD146 could serve as an adhesion molecule active in neural crest cells during embryonic development, highlighting its diverse roles in both normal physiological processes and disease states.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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