MCAM KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 19 bp deletion in exon 6.
HeLa
Human
Cervix
Liquid
Sanger Sequencing
Knockout achieved by using CRISPR/Cas9, Homozygous: 19 bp deletion in exon 6
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
MCAM KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 19 bp deletion in exon 6.
HeLa
Human
Cervix
Liquid
Sanger Sequencing
Knockout achieved by using CRISPR/Cas9, Homozygous: 19 bp deletion in exon 6
Puromycin 1µg/mL
Adenocarcinoma
MCAM
Knockout
CRISPR technology
Sanger Sequencing
Homozygous
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
EU: 2 US: 2
Adherent
Female
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
DMEM (High Glucose) + 10% FBS
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Dry Ice
-196°C
-196°C
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
Recommended control: Human wild-type HeLa cell line (Human wild-type HeLa cell line ab255928). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
We will provide viable cells that proliferate on revival.
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.
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.
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.
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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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 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° 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
Performed under reducing conditions.
Predicted band size: 72 kDa
Observed band size: 120 kDa
Homozygous: 19 bp deletion in exon 6.
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