Human MCAM/CD146 Antibody

Detection of Human MCAM/CD146 by Simple Western^TM.

Simple Western lane view shows lysates of K562 human chronic myelogenous leukemia cell line, loaded at 0.2 mg/mL. A specific band was detected for MCAM/CD146 at approximately 152 kDa (as indicated) using 10 µg/mL of Goat Anti-Human MCAM/CD146 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF932) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

MCAM/CD146 in Human Melanoma.

MCAM/CD146 was detected in immersion fixed paraffin-embedded sections of human melanoma tissue using Goat Anti-Human MCAM/CD146 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF932) at 5 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

MCAM/CD146 in Human Melanoma Tissue.

MCAM/CD146 was detected in immersion fixed paraffin-embedded sections of human melanoma tissue using Goat Anti-Human MCAM/CD146 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF932) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to plasma membrane in cancer cells. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of Human MCAM/CD146 by Immunocytochemistry/ Immunofluorescence

MCAM and COLEC12 are novel BEC- and LEC- specific markers.Two new endothelial markers MCAM and COLEC12 (selected from our analysis of Dataset B) were used to stain (A) normal human lymph nodes together with two established vascular markers PV-1 (for BEC) and CLEVER-1 (for LEC). In addition, (B) chronically inflamed tonsils and (C) specimens from bladder cancer and colorectal cancer were stained with the antibodies against the indicated proteins. LYVE-1 and COLEC12 co-staining was done on colorectal cancer specimens, whereas the other stainings represent bladder cancer. MCAM staining colocalized very well with the established BEC marker PV-1 whereas no colocalization could be detected between MCAM and the established LEC markers LYVE-1 and podoplanin. COLEC12-staining on the other hand showed colocalization with LYVE-1 but not PV-1. White arrows point to areas of colocalization. Nuclear counterstaining was performed with DAPI. Scale bars represent 100 µm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/24058540), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human MCAM/CD146 by Immunocytochemistry/ Immunofluorescence

MCAM and COLEC12 are novel BEC- and LEC- specific markers.Two new endothelial markers MCAM and COLEC12 (selected from our analysis of Dataset B) were used to stain (A) normal human lymph nodes together with two established vascular markers PV-1 (for BEC) and CLEVER-1 (for LEC). In addition, (B) chronically inflamed tonsils and (C) specimens from bladder cancer and colorectal cancer were stained with the antibodies against the indicated proteins. LYVE-1 and COLEC12 co-staining was done on colorectal cancer specimens, whereas the other stainings represent bladder cancer. MCAM staining colocalized very well with the established BEC marker PV-1 whereas no colocalization could be detected between MCAM and the established LEC markers LYVE-1 and podoplanin. COLEC12-staining on the other hand showed colocalization with LYVE-1 but not PV-1. White arrows point to areas of colocalization. Nuclear counterstaining was performed with DAPI. Scale bars represent 100 µm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/24058540), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human MCAM/CD146 by Immunocytochemistry/ Immunofluorescence

MCAM and COLEC12 are novel BEC- and LEC- specific markers.Two new endothelial markers MCAM and COLEC12 (selected from our analysis of Dataset B) were used to stain (A) normal human lymph nodes together with two established vascular markers PV-1 (for BEC) and CLEVER-1 (for LEC). In addition, (B) chronically inflamed tonsils and (C) specimens from bladder cancer and colorectal cancer were stained with the antibodies against the indicated proteins. LYVE-1 and COLEC12 co-staining was done on colorectal cancer specimens, whereas the other stainings represent bladder cancer. MCAM staining colocalized very well with the established BEC marker PV-1 whereas no colocalization could be detected between MCAM and the established LEC markers LYVE-1 and podoplanin. COLEC12-staining on the other hand showed colocalization with LYVE-1 but not PV-1. White arrows point to areas of colocalization. Nuclear counterstaining was performed with DAPI. Scale bars represent 100 µm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/24058540), licensed under a CC-BY license. Not internally tested by R&D Systems.