Mouse MCAM/CD146 Antibody

Catalog # Availability Size / Price Qty
AF6106
AF6106-SP
Detection of Human and Mouse MCAM/CD146 by Western Blot.
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Mouse MCAM/CD146 Antibody Summary

Species Reactivity
Mouse
Specificity
Detects mouse MCAM/CD146 in direct ELISAs and Western blots. In direct ELISAs, approximately 40% cross‑reactivity with recombinant rat MCAM is observed and approximately 10% cross-reactivity with recombinant human MCAM is observed.
Source
Polyclonal Sheep IgG
Purification
Antigen Affinity-purified
Immunogen
Mouse myeloma cell line NS0-derived recombinant mouse MCAM/CD146
Val24-Tyr563
Accession # Q8R2Y2
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.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below
Simple Western
10 µg/mL
See below
Immunocytochemistry
5-15 µg/mL
See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Human and Mouse MCAM/CD146 antibody by Western Blot. View Larger

Detection of Human and Mouse MCAM/CD146 by Western Blot. Western blot shows lysates of Bowes human melanoma cell line and mouse embryo tissue. PVDF Membrane was probed with 1 µg/mL of Sheep Anti-Mouse MCAM/CD146 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6106) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for MCAM/CD146 at approximately 117 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

Immunocytochemistry MCAM/CD146 antibody in B16-F1 Mouse Cell Line by Immunocytochemistry (ICC). View Larger

MCAM/CD146 in B16‑F1 Mouse Cell Line. MCAM/CD146 was detected in immersion fixed B16-F1 mouse melanoma cell line using Sheep Anti-Mouse MCAM/CD146 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6106) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to plasma membranes. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Simple Western Detection of Mouse MCAM/CD146 antibody by Simple Western<sup>TM</sup>. View Larger

Detection of Mouse MCAM/CD146 by Simple WesternTM. Simple Western lane view shows lysates of mouse embryo tissue (15 d.p.c.), loaded at 0.2 mg/mL. A specific band was detected for MCAM/CD146 at approximately 138 kDa (as indicated) using 10 µg/mL of Sheep Anti-Mouse MCAM/CD146 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6106) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Immunocytochemistry/ Immunofluorescence Detection of Mouse MCAM/CD146 by Immunocytochemistry/Immunofluorescence View Larger

Detection of Mouse MCAM/CD146 by Immunocytochemistry/Immunofluorescence Perivascular cells and subaortic mesenchyme are the main source of BMPER within the AGM region. (A) Distribution of BMPER protein in a transverse section of E10.5 AGM measured by immunostaining. Green, CD31; magenta, BMPER; cyan, RUNX1; blue, DAPI. gut, hindgut; nc, notochord. Bar, 50 µm. (B) Bmper mRNA in transverse section of the E10.5 AGM region by in situ hybridization. Bar, 50 µm. (C) Quantification of the mean immunostaining signal intensity (mean gray values) of DAPI (blue) and BMPER (magenta) along a box (not depicted) drawn over the dorsal-ventral axis of the AGM region from A. Distance is from the notochord (dorsal), position 0, to the intersection with the gut and the AGM region (ventral), position 500. (D) Higher magnification of the highlighted region from A showing the aortic endothelium and perivascular population. Green, CD31; magenta, BMPER; cyan, RUNX1; blue, DAPI. Bar, 50 µm. (E) Higher magnification of the region highlighted in B showing Bmper mRNA around the lining of the aorta. Bar, 50 µm. (F) Expression level of Bmper relative to Tbp in each sorted population: Lin−VC−CD45+, representing hematopoietic cells; Lin−VC+CD45−, endothelial cells; Lin−VC−CD45−CD146+, putative perivascular cells; and Lin−VC−CD45−CD146−, remaining stroma. Expression was normalized to the Lin−VC−CD45−CD146+ population. Each population as percentage of Lin− cells indicated below. Sorting was performed twice, first from one pool of embryos from four to five litters and second from two pools of embryos from four to five litters. Error bars represent SD from the mean (n = 3). Significance calculated by t test: **, P = 0.0016; ***, perivascular versus stroma, P = 0.0006; perivascular versus hematopoietic, P = 0.0002. (G) The distribution of nonendothelial, CD146-positive cells and endothelial CD146-positive cells in transverse section of the E10.5 AGM region. Green, CD31; red, CD146; blue, DAPI. Bar, 50 µm. (H) Higher magnification view of the region highlighted in G showing CD31- and CD146-positive endothelial layer and the CD146-positive CD31-negative perivascular layer around the dorsal aorta. Green, CD31; red, CD146; blue, DAPI. Bar, 50 µm. (I) Expression level of Bmper relative to Tbp in each sorted population: Lin−VC+CD45−CD43−CD41lo proHSC, Lin−VC+CD43+CD41+ type I preHSC, and Lin−VC+CD45+ type II preHSC and Lin−VC−CD45−CD146+ putative perivascular cells. Each population as percentage of Lin− cells indicated below. Sorting was performed twice, from pools of two and six litters, respectively. Expression was normalized to the Lin−VC−CD45−CD146+ population. Error bars represent SD from the mean (n = 2). Significance calculated by t test: *, perivascular versus preHSCI, P = 0.04; perivascular versus preHSCII, P = 0.03. (J) Higher magnification view of highlighted region from A showing the localization of BMPER protein in the hematopoietic clusters of the E10.5 dorsal aorta in sections measured by immunostaining. Magenta, BMPER; green, CD31; cyan, RUNX1; blue, DAPI. Bar, 50 µm. (K and L) Higher magnification view of highlighted region from B (K) and from Fig. S3 C (L) showing Bmper mRNA in some but not all cells of the intra-aortic cluster by in situ hybridization. Bars, 50 µm. Image collected and cropped by CiteAb from the following publication (https://rupress.org/jem/article/214/12/3731/42286/A-molecular-roadmap-of-the-AGM-region-reveals), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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.

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
Entrez Gene IDs
4162 (Human); 84004 (Mouse); 78967 (Rat)
Alternate Names
CD146 antigen; CD146; cell surface glycoprotein MUC18; Cell surface glycoprotein P1H12; L-Gicerin; MCAM; melanoma adhesion molecule; melanoma cell adhesion moleculeS-endo 1 endothelial-associated antigen; Melanoma-associated antigen A32; Melanoma-associated antigen MUC18; MUC18; MUC18Gicerin

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