DSG2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1.
ARVC 10, ARVD 10, CDHF 5, CMD1BB, Cadherin family member 5, DSG2_HUMAN, Desmoglein-2, HDGC, HDGC included, Human Desmoglein colon, MGC117034, MGC117036, MGC117037
DSG2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1.
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.
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
Desmoglein 2 (DSG2) is a member of the desmoglein family of proteins sometimes referred to as desmoglein-2 or DSG 2. This protein is a calcium-binding transmembrane glycoprotein and part of the cadherin family with a molecular mass of approximately 122 kDa. DSG2 plays a mechanical role in cell-cell adhesion by forming desmosomes which are specialized structures for maintaining the integrity of tissues. DSG2 is mainly expressed in epithelial cells including those found in the heart skin and certain internal organs.
DSG2 contributes to the structural stability of tissues by locking cells together forming a mechanical and signaling function. The protein is integral to the composition of desmosomes complexes that include other components such as plakoglobin and desmoplakin. These complexes strengthen intercellular adhesion which is essential for tissues that experience constant physical stress like the heart and skin. DSG2 actively participates in cell signaling impacting cellular behavior and communication.
DSG2 plays significant roles in the heart development and epidermal differentiation pathways. DSG2 interacts with several other proteins including plakoglobin and desmoplakin to maintain cellular adhesion and tissue morphogenesis. In the cardiac setting the heart development pathway connects DSG2 with other proteins essential for maintaining the integrity and signaling within the cardiac muscle. This interaction helps modulate pathways that ensure proper cardiac function and repair processes.
DSG2 is linked to arrhythmogenic right ventricular cardiomyopathy (ARVC) which affects the heart's rhythm and muscle structure. DSG2 mutations can disrupt desmosomal structure weakening cell adhesion and leading to disease. Another related disorder is pemphigus vulgaris an autoimmune disease where antibodies target DSG2 among other desmogleins disrupting cell adhesion in the skin and mucous membranes. In ARVC DSG2 frequently connects with other proteins like plakoglobin that are involved in desmosomal interactions leading to the structural and functional deficits seen in the disease.
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Anti-Desmoglein 2/DSG2 antibody [EPR6768] ab150372 was shown to react with Desmoglein 2/DSG2 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261826 (knockout cell lysate Human DSG2 (Desmoglein 2) knockout HeLa cell lysate ab257158) was used. Wild-type HeLa and DSG2 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. Anti-Desmoglein 2/DSG2 antibody [EPR6768] ab150372 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) overnight at 4°C at a 1 in 10000 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-Desmoglein 2/DSG2 antibody [EPR6768] (Anti-Desmoglein 2/DSG2 antibody [EPR6768] ab150372) at 1/10000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: DSG2 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human DSG2 (Desmoglein 2) knockout HeLa cell line (ab261826)
Performed under reducing conditions.
Predicted band size: 122 kDa
Observed band size: 90-160 kDa
Homozygous: 1 bp insertion in exon 1.
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