MCAM/CD146 Knockout HeLa Cell Lysate

Product Discontinued

MCAM/CD146 Knockout HeLa Cell Lysate Summary

• Preparation Method: Knockout achieved by using CRISPR/Cas9,19 bp deletion in exon 6
• Gene: MCAM

Applications/Dilutions

• Dilutions:
  • Western Blot
• Application Notes: You will receive 1 vial (100ug) of knockout cell lysate and 1 vial (100ug) of Parental cell lysate. Lysate can be diluted with 1X SDS sample buffer and will be stable at -20 degrees C for 12 months. Minimize freeze-thaw cycles.

Packaging, Storage & Formulations

• Storage: Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles.
• Buffer: 0.1 mg cell homogenate lyophilized in RIPA buffer made with double-knockout cell lines.
• Concentration: LYOPH
• Reconstitution Instructions: To use as WB negative control, spin down briefly and resuspend in 100 uL 1xSDS sample buffer (2% SDS, 60 mM Tris-HCl pH 6.8, 10% Glycerol, 0.02% Bromophenol blue, 60 mM beta-mercaptoethanol). Boil the lysate for 3 - 5 minutes before loading it onto gel.

Lysate Details for Array

• Type: Knockout HeLa Cell

Notes

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Validation of antibody specificity is critical and verification of antibody performance against knockout samples is one way to guarantee that an antibody recognizes a specific target. Novus' KO cell lysate can be used as a negative control for western blots and to confirm the specificity of antibodies.

Alternate Names for MCAM/CD146 Knockout HeLa Cell Lysate

• CD146 antigen
• CD146
• cell surface glycoprotein MUC18
• Cell surface glycoprotein P1H12
• L-Gicerin
• MCAM
• melanoma adhesion molecule
• melanoma cell adhesion moleculeS-endo 1 endothelial-associated antigen
• Melanoma-associated antigen A32
• Melanoma-associated antigen MUC18
• MUC18
• MUC18Gicerin

Background

CD146 belongs to the immunoglobulin supergene family with five immunoglobulin like domains (V-V-C2-C2-C2), a transmembrane region and a 63 residue cytoplasmic tail. The protein is a membrane glycoprotein that functions as a Ca2+ independent cell adhesion molecule involved in heterophilic cell to cell interactions. CD146 has a molecular size of 130 kDa in its reduced form (118 kDa unreduced), and N linked glycosylation accounts for fifty percent of the apparent molecular weight. In some cells the molecule carries a sulfate 3 glucuronyl moiety. Expression of the molecule was shown in a relatively limited spectrum of normal human tissues (endothelium, smooth muscle, and subpopulations of activated T cells) and malignant neoplasm (melanoma and breast carcinoma). The lineage specific expression pattern of CD146 can be useful in the differential diagnosis of certain lesions including melanomas and various types of gestational trophoblastic lesions. CD146 expression can promote tumor progression in human melanoma, through enhanced interaction between melanoma cells and endothelial cells. However, in breast carcinoma, CD146 may act as a tumor suppressor. Over expression of CD146 in breast carcinoma cells results in a more cohesive cell growth and in the formation of smaller tumors in nude mice. During implantation and placentation, CD146 is expressed by the intermediate trophoblast in the placental site and binds to its putative receptor in uterine smooth muscle cells thus limiting trophoblastic invasion in the myometrium. Monoclonal antibody specific for CD146 is an important tool for the identification and isolation of cells expressing CD146.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Bioinformatics

• Gene Symbol: MCAM