CD146/MCAM Monoclonal antibody, PBS Only

CD146/MCAM Monoclonal Antibody for WB, IF, IHC, Indirect ELISA

Host / Isotype

Mouse / IgG1

Reactivity

human

Applications

WB, IF, IHC, Indirect ELISA

Conjugate

Unconjugated

CloneNo.

4D8A9

Cat no : 66153-1-PBS

Synonyms

CD146, MCAM, MUC18

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Validation Data Gallery

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  WB analysis of A375 using 66153-1-Ig (same clone as 66153-1-PBS)

  A375 cells were subjected to SDS PAGE followed by western blot with 66153-1-Ig (CD146/MCAM antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 66153-1-PBS in a different storage buffer formulation.

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  WB analysis of HepG2 using 66153-1-Ig (same clone as 66153-1-PBS)

  Untreated and PNGase F-treated lysates of HepG2 cells and L02 cells were subjected to SDS PAGE followed by western blot with 66153-1-Ig (CD146/MCAM antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours. PNGase F was obtained from Atagenix (cat.NO. ata808). This data was developed using the same antibody clone with 66153-1-PBS in a different storage buffer formulation.

• 

  WB analysis using 66153-1-Ig (same clone as 66153-1-PBS)

  Various lysates were subjected to SDS PAGE followed by western blot with 66153-1-Ig (CD146/MCAM antibody at dilution of 1:10000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 66153-1-PBS in a different storage buffer formulation.

• 

  WB analysis of HeLa using 66153-1-Ig (same clone as 66153-1-PBS)

  HeLa cells were subjected to SDS PAGE followed by western blot with 66153-1-Ig (CD146/MCAM antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 66153-1-PBS in a different storage buffer formulation.

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  IHC staining of human liver cancer using 66153-1-Ig (same clone as 66153-1-PBS)

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 66153-1-Ig (CD146/MCAM antibody) at dilution of 1:2000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). This data was developed using the same antibody clone with 66153-1-PBS in a different storage buffer formulation.

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  IHC staining of human liver cancer using 66153-1-Ig (same clone as 66153-1-PBS)

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 66153-1-Ig (CD146/MCAM antibody) at dilution of 1:2000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). This data was developed using the same antibody clone with 66153-1-PBS in a different storage buffer formulation.

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  IHC staining of human placenta using 66153-1-Ig (same clone as 66153-1-PBS)

  Immunohistochemical analysis of paraffin-embedded human placenta tissue slide using 66153-1-Ig (CD146/MCAM antibody) at dilution of 1:8000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). This data was developed using the same antibody clone with 66153-1-PBS in a different storage buffer formulation.

• 

  IHC staining of human placenta using 66153-1-Ig (same clone as 66153-1-PBS)

  Immunohistochemical analysis of paraffin-embedded human placenta tissue slide using 66153-1-Ig (CD146/MCAM antibody) at dilution of 1:8000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). This data was developed using the same antibody clone with 66153-1-PBS in a different storage buffer formulation.

• 

  IF Staining of human liver cancer using 66153-1-Ig (same clone as 66153-1-PBS)

  Immunofluorescent analysis of (4% PFA) fixed human liver cancer tissue using CD146/MCAM antibody (66153-1-Ig, Clone: 4D8A9 ) at dilution of 1:2000 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L). This data was developed using the same antibody clone with 66153-1-PBS in a different storage buffer formulation.

• 

  IF Staining of human liver cancer using 66153-1-Ig (same clone as 66153-1-PBS)

  Immunofluorescent analysis of (4% PFA) fixed human liver cancer tissue using CD146/MCAM antibody (66153-1-Ig, Clone: 4D8A9 ) at dilution of 1:2000 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L). This data was developed using the same antibody clone with 66153-1-PBS in a different storage buffer formulation.

• 

  IF Staining of HUVEC using 66153-1-Ig (same clone as 66153-1-PBS)

  Immunofluorescent analysis of (4% PFA) fixed HUVEC cells using CD146/MCAM antibody (66153-1-Ig, Clone: 4D8A9 ) at dilution of 1:500 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L). This data was developed using the same antibody clone with 66153-1-PBS in a different storage buffer formulation.

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Recommended dilution

Application	Dilution
It is recommended that this reagent should be titrated in each testing system to obtain optimal results.
Sample-dependent, Check data in validation data gallery.

Product Information

66153-1-PBS targets CD146/MCAM in WB, IF, IHC, Indirect ELISA applications and shows reactivity with human samples.

• Tested Reactivity: human
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: CD146/MCAM fusion protein Ag11855
• Full Name: melanoma cell adhesion molecule
• Calculated Molecular Weight: 646 aa, 72 kDa
• Observed Molecular Weight: 120 kDa
• GenBank Accession Number: BC056418
• Gene Symbol: MCAM
• Gene ID (NCBI): 4162
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G purification
• Storage Buffer: PBS Only
• Storage Conditions: Store at -80°C.

Background Information

CD146, also known as melanoma cell adhesion molecule (MCAM) or MUC18, originally identified as a biomarker of melanoma progression, is a transmembrane glycoprotein of 113-130 kDa, belonging to the immunoglobulin (Ig) superfamily (PMID: 8378324; 25993332). Structurally, it consists of five Ig domains, a transmembrane domain, and a cytoplasmic region. In normal adult tissue, CD146 is primarily expressed by vascular endothelium and smooth muscle. CD146 is a key cell adhesion protein in vascular endothelial cell activity and angiogenesis, and has been used as marker of circulating endothelium cells (CECs) (PMID: 19356677). In addition to the membrane-anchored form of CD146, a soluble form of CD146 (sCD146, 105 kDa) has also been found in human plasma and in the supernatant of cultured human endothelial cells (PMID: 9462829; 19229070; 16374253; 14597988). This antibody detects a band at approximately 120 kDa that corresponds to the molecular weight of glycosylated CD146. Treatment of lysates of HepG2 cells and L02 cells with PNGase F, which removes oligosaccharides from N-linked glycoproteins, led to a down-shift of the detected band.