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Anti-CD146 antibody [EPR3208] ab75769 is a rabbit monoclonal antibody that is used in CD146 western blotting, IHC, immunofluorescence and flow cytometry. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Antibody clone EPR3208 has been tried and trusted by researchers since 2009 and is cited in >180 publications
- Specificity confirmed with MCAM knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation


Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (AB75769), expandable thumbnail
  • Western blot - Anti-CD146 antibody [EPR3208] (AB75769), expandable thumbnail
  • Western blot - Anti-CD146 antibody [EPR3208] (AB75769), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-CD146 antibody [EPR3208] (AB75769), expandable thumbnail
  • Western blot - Anti-CD146 antibody [EPR3208] (AB75769), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PWBICC/IFFlow Cyt (Intra)IHC-Fr
Human
Tested
Tested
Tested
Tested
Not recommended
Mouse
Expected
Tested
Expected
Expected
Not recommended
Rat
Expected
Tested
Expected
Expected
Not recommended

Tested
Tested

Species

Human

Dilution info

1/250 - 1/500

Notes

Perform heat-mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/1000

Notes

For unpurified use at 1/10000 - 1/50000.

Species

Rat

Dilution info

1/1000

Notes

For unpurified use at 1/10000 - 1/50000.

Species

Human

Dilution info

1/1000

Notes

For unpurified use at 1/10000 - 1/50000.

Tested
Tested

Species

Human

Dilution info

1/100 - 1/250

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

1/20 - 1/80

Notes

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Mouse, Rat, Human

Dilution info

-

Notes

-

Associated Products

Select an associated product type

11 products for Alternative Product

Target data

Function

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 a surface receptor that triggers tyrosine phosphorylation of FYN and PTK2/FAK1, and a transient increase in the intracellular calcium concentration.

Alternative names

MUC18, MCAM, MUC18, Cell surface glycoprotein MUC18, Cell surface glycoprotein P1H12, Melanoma cell adhesion molecule, Melanoma-associated antigen A32, Melanoma-associated antigen MUC18, S-endo 1 endothelial-associated antigen

Recommended products

Anti-CD146 antibody [EPR3208] ab75769 is a rabbit monoclonal antibody that is used in CD146 western blotting, IHC, immunofluorescence and flow cytometry. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Antibody clone EPR3208 has been tried and trusted by researchers since 2009 and is cited in >180 publications
- Specificity confirmed with MCAM knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EPR3208

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

CD146 also known as MCAM or MUC18 is a cell adhesion molecule that belongs to the immunoglobulin superfamily. With a mass of approximately 113 kDa CD146 is expressed mainly on the surface of endothelial cells but it can also be found on smooth muscle cells some T-cells and in certain cancerous tissues. This protein plays a role in cell-cell adhesion contributing to the integrity and signaling functions within tissue microenvironments.

Biological function summary

CD146 influences the migration and organization of cells within the vascular system. It serves as a component of cell adhesion complexes facilitating interactions between endothelial cells and other cell types. Additionally CD146 contributes to angiogenesis the process by which new blood vessels form from existing vasculature which is critical during tissue development and repair. This function makes it an important element in maintaining normal physiological processes.

Pathways

CD146 participates in the Wnt/β-catenin and MAPK signaling pathways which are vital in cell proliferation and differentiation. Through the Wnt/β-catenin pathway CD146 interacts with other proteins like Frizzled receptors to regulate gene transcription. In the MAPK pathway it is linked with signaling cascades that involve proteins such as ERK leading to cellular responses necessary for growth and response to external stimuli.

Associated diseases and disorders

CD146 has been implicated in melanoma progression and cardiovascular diseases. Its overexpression in melanoma associates it with increased tumor growth and metastasis often involving interactions with other adhesion molecules like integrins. In cardiovascular diseases CD146 relates to atherosclerosis as its expression on endothelial cells can promote inflammatory responses. Understanding these connections provides insights into potential therapeutic targets for treating these conditions.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

17 product images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769), expandable thumbnail
    Image from Davaine J.M. et al. PLoS One. 2014 Sep 26;9(9):e107642. doi: 10.1371/journal.pone.0107642. eCollection 2014.

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769)

    Immunohistochemistry experiments were used to compare symptomatic carotid plaques (SC) and asymptomatic carotid plaques (AsC)

    Asymptomatic lesions presented higher CD146+ pericyte infiltration, p<0.001. Representative images are on the left with corresponding quantification on the right.

    ab75769 used at 1/200 dilution.

    (After Figure 2 of Davaine et al)

  • Western blot - Anti-CD146 antibody [EPR3208] (ab75769), expandable thumbnail

    Western blot - Anti-CD146 antibody [EPR3208] (ab75769)

    Lanes 1- 2: Merged signal (red and green). Green - ab75769 observed at 120 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.

    ab75769 was shown to react with CD146 in wild-type HeLa cells in western blot. The band observed in knockout cell line Human MCAM (CD146) knockout HeLa cell line ab261790 (knockout cell lysate Human MCAM (CD146) knockout HeLa cell lysate ab256985) lane below 120kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and MCAM knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab75769 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-CD146 antibody [EPR3208] (ab75769) at 1/1000 dilution

    Lane 1: Wild-type HeLa cell lysate at 20 µg

    Lane 2: MCAM knockout HeLa cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 72 kDa

    Observed band size: 120 kDa

  • Western blot - Anti-CD146 antibody [EPR3208] (ab75769), expandable thumbnail

    Western blot - Anti-CD146 antibody [EPR3208] (ab75769)

    Lane 1: Wild-type HAP1 whole cell lysate (40 μg)
    Lane 2: CD146 knockout HAP1 whole cell lysate (40 μg)
    Lane 3: A375 whole cell lysate (20 μg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab75769 observed at 120-72 kDa. Red - loading control, Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484, observed at 37 kDa.

    ab75769 was shown to specifically react with CD146 in wild-type HAP1 cells as signal was lost in CD146 knockout cells. Wild-type and CD146 knockout samples were subjected to SDS-PAGE. ab75769 and Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/10000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-CD146 antibody [EPR3208] (ab75769)

    Predicted band size: 72 kDa

  • Flow Cytometry (Intracellular) - Anti-CD146 antibody [EPR3208] (ab75769), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-CD146 antibody [EPR3208] (ab75769)

    Intracellular Flow Cytometry analysis of A375 (human malignant melanoma) cells labeling CD146 with unpurified ab75769 at 1/20 dilution (10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluorr®488) at 1/2000 dilution was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

  • Western blot - Anti-CD146 antibody [EPR3208] (ab75769), expandable thumbnail

    Western blot - Anti-CD146 antibody [EPR3208] (ab75769)

    Lanes 1- 2: Merged signal (red and green). Green - ab75769 observed at 120 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.

    ab75769 was shown to react with CD146 in wild-type HeLa cells in western blot. The band observed in CRISPR/Cas9 edited cell line Human MCAM (CD146) knockout HeLa cell line ab261790 (CRISPR/Cas9 edited cell lysate Human MCAM (CD146) knockout HeLa cell lysate ab256985) lane below 120kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and MCAM CRISPR/Cas9 edited HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab75769 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-CD146 antibody [EPR3208] (ab75769) at 1/1000 dilution

    Lane 1: Wild-type HeLa cell lysate at 20 µg

    Lane 2: MCAM CRISPR/Cas9 edited HeLa cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 72 kDa

    Observed band size: 120 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labelling CD146 with purified ab75769 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) ab97051, a goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Western blot - Anti-CD146 antibody [EPR3208] (ab75769), expandable thumbnail

    Western blot - Anti-CD146 antibody [EPR3208] (ab75769)

    Blocking buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM /TBST.

    All lanes: Western blot - Anti-CD146 antibody [EPR3208] (ab75769) at 1/10000 dilution

    Lane 1: A375 cell lysate at 20 µg

    Lane 2: Human fetal artery lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/1000 dilution

    Predicted band size: 72 kDa

  • Western blot - Anti-CD146 antibody [EPR3208] (ab75769), expandable thumbnail

    Western blot - Anti-CD146 antibody [EPR3208] (ab75769)

    Blocking buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM /TBST.

    All lanes: Western blot - Anti-CD146 antibody [EPR3208] (ab75769) at 1/10000 dilution

    All lanes: HUVEC cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/1000 dilution

    Predicted band size: 72 kDa

  • Flow Cytometry (Intracellular) - Anti-CD146 antibody [EPR3208] (ab75769), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-CD146 antibody [EPR3208] (ab75769)

    Intracellular Flow Cytometry analysis ofHUVEC cells labelling CD146 with purified ab75769 at 1/50 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • Western blot - Anti-CD146 antibody [EPR3208] (ab75769), expandable thumbnail

    Western blot - Anti-CD146 antibody [EPR3208] (ab75769)

    Blocking buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM /TBST.

    All lanes: Western blot - Anti-CD146 antibody [EPR3208] (ab75769) at 1/10000 dilution

    All lanes: B16-F0 cell lysate at 20 µg/mL

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/1000 dilution

    Predicted band size: 72 kDa

  • Western blot - Anti-CD146 antibody [EPR3208] (ab75769), expandable thumbnail

    Western blot - Anti-CD146 antibody [EPR3208] (ab75769)

    Blocking buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM /TBST.

    All lanes: Western blot - Anti-CD146 antibody [EPR3208] (ab75769) at 1/10000 dilution

    All lanes: Rat placenta lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/1000 dilution

    Predicted band size: 72 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of melanoma tissue labelling CD146 with unpurified ab75769 at 1/250. A HRP/AP polymerized secondary antibody was used.

    Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of breast carcinoma vessels tissue labelling CD146 with unpurified ab75769.

    Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of urinary bladder transitional carcinoma vessels tissue labelling CD146 with unpurified ab75769.

    Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of glioma vessels tissue labelling CD146 with unpurified ab75769.

    Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal tonsil tissue labelling CD146 with unpurified ab75769.

    Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (ab75769)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal spleen tissue labelling CD146 with unpurified ab75769.

    Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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