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Immunofluorescence staining of frozen sections of human kidney showing distribution of CD146 (Catalog Number MAB16985 clone P1H12) in endothelium of b...
Western Blot Analysis:
HUVEC lysate was resolved by electrophoresis, transferred to PVDF membrane and probed with anti-MCAM (1:500 dilution of th...
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Anti-MCAM, clone P1H12
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| Ca, H, M, Rb, not R | ELISA, FC, IC, IH, IP, WB, not IH(P) | Mouse | Purified | Monoclonal Antibody |
Description:
Anti-MCAM, clone P1H12
Trade Name:
Chemicon (Millipore)
Specificity:
P1H12 reacts specifically with CD146 {MCAM, MUC18}. In blood and marrow P1H12 reacts only with endothelial cells, and has been used to detect circulating endothelial cells in human, dogs, rabbits and mouse samples. It 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, or EBV-transformed B cell lines, although it expected that CD146 expression is present in many tumor lines. P1H12 does not stain 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 P1H12 also are 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.
Molecular Weight:
110 kDa
Immunogen:
Immunization of mice with HUVEC.
Clone:
P1H12
Isotype:
IgG1
Background Information:
MUC18 antigen (CD146), a member of the immuoglobulin superfamily, is an intrinsic membrane glycoprotein of 110-120 kDa found on the surface of endothelial cells, bone marrow fibroblasts and various melanomas. MUC18 has been used as a marker of tumor progression in human melanoma because expression in those tumors correlates strongly with poor prognosis and the development of metastic disease. In addition, a number of human T, B and myeloid leukemic cell lines seem to express MUC18. The close structural relationship with N-CAM and related molecules suggests that MUC18 may be also a developmentally regulated cell adhesion molecule (Melanoma adhesion molecule or MCAM).
Species Reactivity:
- Canine
- Human
- Mouse
- Rabbit
Species Reactivity Note:
Detects circulating endothelial cells in human. Expected to cross-react with dog, rabbit & mouse.
Non-Reactive Species:
Rat
Application Notes:
Immunocytochemistry:
1-10 μg/mL of a previous lot worked in immunocytochemistry.Works best on EDTA or Trypsin lifted endothelial cells.
Immunohistochemistry:
1-10 µg/mL. 4% PFA for 30min RT or <2hrs at 4°C. Block w/ 1%BSA/0.2% tween20/PBS for 30min. Works well in frozen tissue; fixed or unfixed.
Immunoprecipitation:
1-10 μg/mL of a previous lot worked in immunoprecipitation.
ELISA:
1-10 μg/mL of a previous lot worked in ELISA.
FACS Analysis:
1-10 μg/mL of a previous lot worked in FACS.
Optimal working dilutions must be determined by end user.
1-10 μg/mL of a previous lot worked in immunocytochemistry.Works best on EDTA or Trypsin lifted endothelial cells.
Immunohistochemistry:
1-10 µg/mL. 4% PFA for 30min RT or <2hrs at 4°C. Block w/ 1%BSA/0.2% tween20/PBS for 30min. Works well in frozen tissue; fixed or unfixed.
Immunoprecipitation:
1-10 μg/mL of a previous lot worked in immunoprecipitation.
ELISA:
1-10 μg/mL of a previous lot worked in ELISA.
FACS Analysis:
1-10 μg/mL of a previous lot worked in FACS.
Optimal working dilutions must be determined by end user.
Applications Not Recommended:
Immunohistochemistry (Paraffin)
Control:
HUVEC cells.
Quality Assurance:
Routinely evaluated by Western Blot on HUVEC lysates.
Western Blot Analysis:
1:500 dilution of this lot detected MCAM on 10μg of HUVEC lysates.
Western Blot Analysis:
1:500 dilution of this lot detected MCAM on 10μg of HUVEC lysates.
Purification Method:
Protein A purfied
Presentation:
Purified mouse monoclonal IgG1 in buffer containing 0.02 M PB with 0.25 M NaCl, pH=7.6, with 0.1% sodium azide.
Storage Conditions:
Stable for 1 year at 2-8°C from date of receipt.
UniProt Number:
Entrez Gene Number:
Gene Symbol:
- MCAM
- Gicerin
- MUC18
- CD146
Alternate Names:
- CD146 antigen
- Cell surface glycoprotein P1H12
- Melanoma-associated antigen A32
- Melanoma-associated antigen MUC18
- S-endo 1 endothelial-associated antigen
- melanoma adhesion molecule
- melanoma cell adhesion molecule
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Key Applications:
- ELISA
- Flow Cytometry
- Immunocytochemistry
- Immunohistochemistry
- Immunoprecipitation
- Western Blotting
UniProt Summary:
FUNCTION: SwissProt: P43121 # 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.
SIZE: 646 amino acids; 71607 Da
SUBCELLULAR LOCATION: Membrane; Single-pass type I membrane protein.
TISSUE SPECIFICITY: 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.
SIMILARITY: SwissProt: P43121 ## Contains 3 Ig-like C2-type (immunoglobulin-like) domains. & Contains 2 Ig-like V-type (immunoglobulin-like) domains.
SIZE: 646 amino acids; 71607 Da
SUBCELLULAR LOCATION: Membrane; Single-pass type I membrane protein.
TISSUE SPECIFICITY: 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.
SIMILARITY: SwissProt: P43121 ## Contains 3 Ig-like C2-type (immunoglobulin-like) domains. & Contains 2 Ig-like V-type (immunoglobulin-like) domains.
Brand Family:
Chemicon
Product Name:
Anti-MCAM, clone P1H12
Concentration:
1 mg/mL
Antibody Type:
Monoclonal Antibody
Qty/Pk:
100 µg
Format:
Purified
Host:
Mouse