Key Product Details

Validated by

Knockout/Knockdown, Biological Validation

Species Reactivity

Validated:

Mouse

Cited:

Mouse

Applications

Validated:

Flow Cytometry, Immunocytochemistry, CyTOF-ready

Cited:

Immunohistochemistry, Flow Cytometry, Immunocytochemistry

Label

Unconjugated

Antibody Source

Monoclonal Rat IgG2A Clone # 733216
View all MCAM/CD146 Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant mouse MCAM/CD146
Met1-Val563
Accession # Q8R2Y2

Specificity

Detects mouse MCAM/CD146 in direct ELISAs. In direct ELISAs, less than 5% cross-reactivity with recombinant mouse (rm) MAdCAM-1 is observed, and no cross-reactivity with rmALCAM, rmNCAM, rmL1CAM, rmOCAM, rmTROP-2, recombinant human MCAM, or recombinant rat MCAM is observed.

Clonality

Monoclonal

Host

Rat

Isotype

IgG2A

Scientific Data Images for Mouse MCAM/CD146 Antibody

Detection of MCAM/CD146 antibody in NK1.1+ Mouse Splenocytes antibody by Flow Cytometry.

Detection of MCAM/CD146 in NK1.1+ Mouse Splenocytes by Flow Cytometry.

Mouse NK1.1+ splenocytes were stained with Rat Anti-Mouse MCAM/CD146 Monoclonal Antibody (Catalog # MAB7718, filled histogram) or isotype control antibody (Catalog # MAB006, open histogram), followed by Allophycocyanin-conjugated Anti-Rat IgG Secondary Antibody (Catalog # F0113).
MCAM/CD146 antibody in bEnd by Immunocytochemistry (ICC).3 Mouse Cell Line by Immunocytochemistry (ICC).

MCAM/CD146 in bEnd.3 Mouse Cell Line.

MCAM/CD146 was detected in immersion fixed bEnd.3 mouse endothelioma cell line using Rat Anti-Mouse MCAM/CD146 Monoclonal Antibody (Catalog # MAB7718) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the Northern-Lights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red; Catalog # NL013) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces and cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Mouse MCAM/CD146 by Immunocytochemistry/Immunofluorescence

Detection of Mouse MCAM/CD146 by Immunocytochemistry/Immunofluorescence

MCAM is required to establish cell autonomous polarity. (A) In elongating myotubes (10T1/2 cells treated with testosterone for 7 days) VANGL2 is localized asymmetrically at the tip of the cell. (B) The VANGL2 enriched tip of the cell is marked by MSN. (C) In MCAM knockout C164 cells myotube elongation fails, MSN labels the whole plasma membrane and VANGL2 is spread across the cytoplasm. (D) Highly polarized localization of MCAM and SCRIB at the distal end of growing wild-type myotube. Separate channels of the boxed area are shown on the right. (E) In MCAM knockout cells SCRIB levels remain low and it is spread evenly in the cell. (F) In wild-type myotubes PAR3 remains cytoplasmic, whereas (G) in MCAM knockout C164 cells it can be detected at the cell cortex. (H) RT-qPCR demonstrates reduced expression of Scrib in MCAM mutant cell lines. Cells were treated for 7 days with BMP2 or testosterone (n=3; **P<0.01; ***P<0.001; two-tailed t-test; mean±s.e.m.). (I) Deletion of MCAM endocytosis motif leads to similar polarity defects as complete MCAM elimination. VANGL2 is evenly spread in U125 cells and PAR3 accumulates in cell cortex. (J) In chondrogenic differentiation VANGL2 was observed asymmetrically in limited number of cells. In MCAM mutant cell lines (C149, C164, U125) VANGL2 accumulated around the nucleus. (K) Initiation of myogenic (4-day culture with testosterone) and chondrogenic differentiation (4-day culture with BMP2) led to downregulation of ERK1/2 phosphorylation (p-ERK1/2). Instead in MCAM mutant cell lines ERK1/2 phosphorylation increased. Scale bars: 25 µm. Image collected and cropped by CiteAb from the following publication (https://journals.biologists.com/bio/article/doi/10.1242/bio.027771/2567…), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse MCAM/CD146 by Immunocytochemistry/Immunofluorescence

Detection of Mouse MCAM/CD146 by Immunocytochemistry/Immunofluorescence

MCAM is required to establish cell autonomous polarity. (A) In elongating myotubes (10T1/2 cells treated with testosterone for 7 days) VANGL2 is localized asymmetrically at the tip of the cell. (B) The VANGL2 enriched tip of the cell is marked by MSN. (C) In MCAM knockout C164 cells myotube elongation fails, MSN labels the whole plasma membrane and VANGL2 is spread across the cytoplasm. (D) Highly polarized localization of MCAM and SCRIB at the distal end of growing wild-type myotube. Separate channels of the boxed area are shown on the right. (E) In MCAM knockout cells SCRIB levels remain low and it is spread evenly in the cell. (F) In wild-type myotubes PAR3 remains cytoplasmic, whereas (G) in MCAM knockout C164 cells it can be detected at the cell cortex. (H) RT-qPCR demonstrates reduced expression of Scrib in MCAM mutant cell lines. Cells were treated for 7 days with BMP2 or testosterone (n=3; **P<0.01; ***P<0.001; two-tailed t-test; mean±s.e.m.). (I) Deletion of MCAM endocytosis motif leads to similar polarity defects as complete MCAM elimination. VANGL2 is evenly spread in U125 cells and PAR3 accumulates in cell cortex. (J) In chondrogenic differentiation VANGL2 was observed asymmetrically in limited number of cells. In MCAM mutant cell lines (C149, C164, U125) VANGL2 accumulated around the nucleus. (K) Initiation of myogenic (4-day culture with testosterone) and chondrogenic differentiation (4-day culture with BMP2) led to downregulation of ERK1/2 phosphorylation (p-ERK1/2). Instead in MCAM mutant cell lines ERK1/2 phosphorylation increased. Scale bars: 25 µm. Image collected and cropped by CiteAb from the following publication (https://journals.biologists.com/bio/article/doi/10.1242/bio.027771/2567…), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse MCAM/CD146 by Immunocytochemistry/Immunofluorescence

Detection of Mouse MCAM/CD146 by Immunocytochemistry/Immunofluorescence

MCAM is required to establish cell autonomous polarity. (A) In elongating myotubes (10T1/2 cells treated with testosterone for 7 days) VANGL2 is localized asymmetrically at the tip of the cell. (B) The VANGL2 enriched tip of the cell is marked by MSN. (C) In MCAM knockout C164 cells myotube elongation fails, MSN labels the whole plasma membrane and VANGL2 is spread across the cytoplasm. (D) Highly polarized localization of MCAM and SCRIB at the distal end of growing wild-type myotube. Separate channels of the boxed area are shown on the right. (E) In MCAM knockout cells SCRIB levels remain low and it is spread evenly in the cell. (F) In wild-type myotubes PAR3 remains cytoplasmic, whereas (G) in MCAM knockout C164 cells it can be detected at the cell cortex. (H) RT-qPCR demonstrates reduced expression of Scrib in MCAM mutant cell lines. Cells were treated for 7 days with BMP2 or testosterone (n=3; **P<0.01; ***P<0.001; two-tailed t-test; mean±s.e.m.). (I) Deletion of MCAM endocytosis motif leads to similar polarity defects as complete MCAM elimination. VANGL2 is evenly spread in U125 cells and PAR3 accumulates in cell cortex. (J) In chondrogenic differentiation VANGL2 was observed asymmetrically in limited number of cells. In MCAM mutant cell lines (C149, C164, U125) VANGL2 accumulated around the nucleus. (K) Initiation of myogenic (4-day culture with testosterone) and chondrogenic differentiation (4-day culture with BMP2) led to downregulation of ERK1/2 phosphorylation (p-ERK1/2). Instead in MCAM mutant cell lines ERK1/2 phosphorylation increased. Scale bars: 25 µm. Image collected and cropped by CiteAb from the following publication (https://journals.biologists.com/bio/article/doi/10.1242/bio.027771/2567…), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse MCAM/CD146 by Immunocytochemistry/Immunofluorescence

Detection of Mouse MCAM/CD146 by Immunocytochemistry/Immunofluorescence

MCAM is required to establish cell autonomous polarity. (A) In elongating myotubes (10T1/2 cells treated with testosterone for 7 days) VANGL2 is localized asymmetrically at the tip of the cell. (B) The VANGL2 enriched tip of the cell is marked by MSN. (C) In MCAM knockout C164 cells myotube elongation fails, MSN labels the whole plasma membrane and VANGL2 is spread across the cytoplasm. (D) Highly polarized localization of MCAM and SCRIB at the distal end of growing wild-type myotube. Separate channels of the boxed area are shown on the right. (E) In MCAM knockout cells SCRIB levels remain low and it is spread evenly in the cell. (F) In wild-type myotubes PAR3 remains cytoplasmic, whereas (G) in MCAM knockout C164 cells it can be detected at the cell cortex. (H) RT-qPCR demonstrates reduced expression of Scrib in MCAM mutant cell lines. Cells were treated for 7 days with BMP2 or testosterone (n=3; **P<0.01; ***P<0.001; two-tailed t-test; mean±s.e.m.). (I) Deletion of MCAM endocytosis motif leads to similar polarity defects as complete MCAM elimination. VANGL2 is evenly spread in U125 cells and PAR3 accumulates in cell cortex. (J) In chondrogenic differentiation VANGL2 was observed asymmetrically in limited number of cells. In MCAM mutant cell lines (C149, C164, U125) VANGL2 accumulated around the nucleus. (K) Initiation of myogenic (4-day culture with testosterone) and chondrogenic differentiation (4-day culture with BMP2) led to downregulation of ERK1/2 phosphorylation (p-ERK1/2). Instead in MCAM mutant cell lines ERK1/2 phosphorylation increased. Scale bars: 25 µm. Image collected and cropped by CiteAb from the following publication (https://journals.biologists.com/bio/article/doi/10.1242/bio.027771/2567…), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Mouse MCAM/CD146 Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

2.5 µg/106 cells
Sample: Mouse splenocytes

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed mouse splenocytes

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: MCAM/CD146

MCAM (Melanoma cell adhesion molecule; also CD146 and MUC18) is a 110‑120 kDa member of a small group of Ig‑superfamily molecules that includes CD239 and CD166. MCAM has also been reported at a molecular weight of approximately 150 kDa. In rodent, MCAM is reportedly expressed on neurons, endothelial cells, NK cells, neutrophils, mesenchymal stem cells and melanoma cells. MCAM appears to contribute to intercellular endothelial cell junctions, and possibly contributes to the migration of select cell types. Mature mouse MCAM is a 625 amino acid (aa) type I transmembrane glycoprotein. Its extracellular region is 540 aa in length (aa 24‑563). It contains two V‑type Ig‑like domains (aa 24‑244) followed by three C2‑type Ig‑like domains (aa 246‑512). One cytoplasmic region splice form shows a seven aa substitution for aa 600‑648. Unlike human, rodent MCAM does not undergo a splicing event that will generate a soluble isoform. Over aa 24‑563, mouse MCAM shares 90% and 74% aa identity with rat and human MCAM, respectively.

Long Name

Melanoma Cell Adhesion Molecule

Alternate Names

CD146, L-Gicerin, MUC18

Entrez Gene IDs

4162 (Human); 84004 (Mouse); 78967 (Rat)

Gene Symbol

MCAM

UniProt

Q8R2Y2

Additional MCAM/CD146 Products

Product Documents for Mouse MCAM/CD146 Antibody

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Product Specific Notices for Mouse MCAM/CD146 Antibody

For research use only

Citations for Mouse MCAM/CD146 Antibody

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Protocols

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