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AB256985

Human MCAM (CD146) knockout HeLa cell lysate

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MCAM KO cell lysate available now. KO validated by. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 19 bp deletion in exon 6.

View Alternative Names

A32 antigen, CD146 antigen, Cell surface glycoprotein MUC18, Cell surface glycoprotein P1H12, Gicerin, MUC18_HUMAN, Mcam, MelCAM, Melanoma adhesion molecule, Melanoma associated glycoprotein MUC18, Melanoma cell adhesion molecule, Melanoma-associated antigen A32, Melanoma-associated antigen MUC18, S endo 1, S-endo 1 endothelial-associated antigen

2 Images
Western blot - Human MCAM (CD146) knockout HeLa cell lysate (AB256985)
  • WB

Lab

Western blot - Human MCAM (CD146) knockout HeLa cell lysate (AB256985)

Lane 1 : Wild-type HeLa cell lysate (20µg)

Lane 2 : MCAM knockout HeLa cell lysate (20µg)

Lanes 1- 2 : Merged signal (red and green). Green - ab75769 observed at 120 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab75769 Anti-CD146 antibody [EPR3208] was shown to specifically react with CD146 in wild-type HeLa cells in western blot. The band observed in the knockout cell line ab261790 (knockout cell lysate ab256985) lane below 120kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type and CD146 knockout samples 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 (ab8245) were incubated overnight at 4°C at 1 in 1000 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and 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] (<a href='/en-us/products/primary-antibodies/cd146-antibody-epr3208-ab75769'>ab75769</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

MCAM knockout HeLa cell lysate at 20 µg

Predicted band size: 72 kDa

Observed band size: 120 kDa

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Sanger Sequencing - Human MCAM (CD146) knockout HeLa cell lysate (AB256985)
  • Sanger seq

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Sanger Sequencing - Human MCAM (CD146) knockout HeLa cell lysate (AB256985)

Homozygous : 19 bp deletion in exon 6

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 19 bp deletion in exon 6.

Disease

Adenocarcinoma

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p>Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.</p>" } } }
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Product details

Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.

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What's included?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab256985-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human MCAM knockout HeLa cell lysate", "number":"AB256985-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HeLa cell lysate", "number":"AB256985-CMP02", "productcode":"" } ] } } }
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Properties and storage information

Gene name
MCAM
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
Zygosity
Homozygous
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

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

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.

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.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Complementary Products

Primary Antibodies

AB75769

Anti-CD146 antibody [EPR3208]

RabMAb|Recombinant|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [EPR3208] (AB75769)

  • 4
  • 41
Secondary Antibodies

AB150077

Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)

Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (AB150077)

  • 5
  • 20
Cell Lines & Lysates

AB261790

Human MCAM (CD146) knockout HeLa cell line

Advanced Validation

Western blot - Human MCAM (CD146) knockout HeLa cell line (AB261790)

  • 0
  • 0
Primary Antibodies

AB137869

Anti-Tyrosine Hydroxylase antibody [EP1532Y] - Neuronal Marker

RabMAb|Recombinant|Lab Essentials|20ul selling size

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tyrosine Hydroxylase antibody [EP1532Y] - Neuronal Marker (AB137869)

  • 5
  • 9

Product promise

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For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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