Anti-CD146 antibody [EPR3208] - Carrier free (ab210072)

Anti-CD146 [EPR3208] antibody (ab210072) is a rabbit monoclonal antibody that is provided in a PBS only formulation free from BSA and azide. Formulated to be conjugation-ready. It is used to detect CD146 in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IHC-P, ICC/IF. Suitable for Human, Mouse, Rat samples.

- Specificity confirmed with CD146 knockout cell line validation
-BSA, sodium azide, and glycerol-free for consistent conjugation with fluorochromes, enzymes and more

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Related conjugates and formulations

ab75769
Unconjugated
Anti-CD146 antibody [EPR3208]
ab216361
Conjugated
PerCP Anti-CD146 antibody [EPR3208]
ab217072
Conjugated
Biotin Anti-CD146 antibody [EPR3208]
ab209298
Conjugated
PE Anti-CD146 antibody [EPR3208]
ab196448
Conjugated
Alexa Fluor® 488 Anti-CD146 antibody [EPR3208]
ab303023
Conjugated
APC Anti-CD146 antibody [EPR3208]
ab305213
Conjugated
Alexa Fluor® 647 Anti-CD146 antibody [EPR3208]
ab310672
Conjugated
Alexa Fluor® 594 Anti-CD146 antibody [EPR3208]
ab303024
Conjugated
HRP Anti-CD146 antibody [EPR3208]
ab307390
Conjugated
Alexa Fluor® 555 Anti-CD146 antibody [EPR3208]

Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] - BSA and Azide free (AB210072), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
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-P	WB	ICC/IF	Flow Cyt (Intra)
Human	Tested	Tested	Tested	Tested
Mouse	Expected	Tested	Expected	Expected
Rat	Expected	Tested	Expected	Expected

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
Species Rat	Dilution info -	Notes Perform heat-mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Human, Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat, Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Associated Products

Select an associated product type

11 products for Alternative Product

9 products for Alternative Version

Target data

See full target information MCAM

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

CD146, MUC18, MCAM, 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

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
Clone number: EPR3208
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: +4°C
Storage information: Do Not Freeze

Notes

What is this antibody validated in?

Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Flow Cytometry (Intra), Flow Cytometry (Flow Cyt), Immunohistochemistry (IHC-P), Immunocytochemistry/immunofluorescence (ICC/IF) in Human, Mouse, Rat samples.

What is the molecular weight of CD146?

Anti-CD146 [EPR3208] - BSA and Azide free (ab210072) specifically detects a band for CD146 (UniProt: P43121) at a molecular weight of 72kDa.

Trusted by the scientific community

Anti-CD146 [EPR3208] - BSA and Azide free (ab210072) was first used in a scientific publication in 2015 and has been cited over 10 times in peer-reviewed journals.

Trial sizes available!

Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies.

Specificity confirmed

The specificity of Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072) has been confirmed by Western blot testing in MCAM Knockout HAP1 cells.

Other related products

We have a range of other formats of antibody clone [EPR3208] also available for your convenience:
ab75769, Alexa Fluor® 488 - ab196448, PE - ab209298, Carrier free - ab210072, PerCP - ab216361, Biotin - ab217072, APC - ab303023, HRP - ab303024, Alexa Fluor® 647 - ab305213, Alexa Fluor® 555 - ab307390, Alkaline Phosphatase - ab308927, Alexa Fluor® 594 - ab310672

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

What are the advantages of a recombinant monoclonal antibody?
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.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

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:
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12 product images

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] - BSA and Azide free (ab210072)
Immunohistochemistry experiments were used to compare symptomatic carotid plaques (SC) and asymptomatic carotid plaques (AsC)
Asymptomatic lesions presented higher CD146⁺ pericyte infiltration, p
Anti-CD146 antibody [EPR3208] ab75769 used at 1/200 dilution.
(After Figure 2 of Davaine et al)
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD146 antibody [EPR3208] ab75769).
Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
Western blot - Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072)
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 - Anti-CD146 antibody [EPR3208] ab75769 observed at 120-72 kDa. Red - loading control, Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484, observed at 37 kDa.
Anti-CD146 antibody [EPR3208] 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. Anti-CD146 antibody [EPR3208] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD146 antibody [EPR3208] ab75769).
All lanes: Western blot - Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072)
Predicted band size: 72 kDa
Flow Cytometry (Intracellular) - Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072)
Intracellular Flow Cytometry analysis of A375 (human malignant melanoma) cells labeling CD146 with unpurified Anti-CD146 antibody [EPR3208] 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 Fluor^®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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD146 antibody [EPR3208] ab75769).
Western blot - Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072)
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD146 antibody [EPR3208] ab75769).
Lanes 1- 2: Merged signal (red and green). Green - Anti-CD146 antibody [EPR3208] ab75769 observed at 120 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.
Anti-CD146 antibody [EPR3208] 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. Anti-CD146 antibody [EPR3208] 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] (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
Lane 2: Western blot - Human MCAM (CD146) knockout HeLa cell line (Human MCAM (CD146) knockout HeLa cell line ab261790)
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] - BSA and Azide free (ab210072)
This IHC data was generated using the same anti-CD146 antibody clone, EPR3208, in a different buffer formulation (cat# Anti-CD146 antibody [EPR3208] ab75769).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labelling CD146 with purified Anti-CD146 antibody [EPR3208] 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.
Flow Cytometry (Intracellular) - Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072)
Intracellular Flow Cytometry analysis of HUVEC cells labelling CD146 with purified Anti-CD146 antibody [EPR3208] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD146 antibody [EPR3208] ab75769).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] - BSA and Azide free (ab210072)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of melanoma tissue labelling CD146 with unpurified Anti-CD146 antibody [EPR3208] ab75769 at 1/250. A HRP/AP polymerized secondary antibody was used.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD146 antibody [EPR3208] 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] - BSA and Azide free (ab210072)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of breast carcinoma vessels tissue labelling CD146 with unpurified Anti-CD146 antibody [EPR3208] ab75769.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD146 antibody [EPR3208] 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] - BSA and Azide free (ab210072)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of urinary bladder transitional carcinoma vessels tissue labelling CD146 with unpurified Anti-CD146 antibody [EPR3208] ab75769.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD146 antibody [EPR3208] 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] - BSA and Azide free (ab210072)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of glioma vessels tissue labelling CD146 with unpurified Anti-CD146 antibody [EPR3208] ab75769.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD146 antibody [EPR3208] 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] - BSA and Azide free (ab210072)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal tonsil tissue labelling CD146 with unpurified Anti-CD146 antibody [EPR3208] ab75769.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD146 antibody [EPR3208] 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] - BSA and Azide free (ab210072)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal spleen tissue labelling CD146 with unpurified Anti-CD146 antibody [EPR3208] ab75769.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD146 antibody [EPR3208] ab75769).
Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.