OBSCN KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2.
BC046431, Gm878, KIAA1556, KIAA1639, OBSCN_HUMAN, OTTMUSG00000005786, Obscurin, Obscurin-MLCK, Obscurin-RhoGEF, Obscurin-myosin light chain kinase, UNC89
OBSCN KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2.
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.
The OBSCN gene encodes the protein obscurin also known by its alternate names including UNC-89. Obscurin is a large protein with a molecular mass exceeding 800 kDa. It is primarily located in striated muscle where it plays a role in linking the sarcomere to the sarcoplasmic reticulum. Obscurin contains multiple domains that facilitate protein-protein interactions allowing it to interact with various components within muscle cells.
Obscurin assists in maintaining the structural integrity and organization of myofibrils. It forms part of a complex in muscle cells involving proteins such as titin and myomesin. This complex aids in the mechanical stability of the contractile apparatus ensuring muscle contraction occurs effectively. Obscurin’s interactions within this complex contribute to the proper transmission of force across the muscle fiber.
The functions of obscurin link it to the sarcomeric signaling pathway and calcium handling pathways in muscle physiology. These pathways involve coordination with titin and ankyrins which help regulate muscle contraction and relaxation. Through these pathways obscurin influences muscle cell responsiveness to mechanical stress and contributes to muscle homeostasis.
Altered obscurin function associates with muscle-related conditions particularly hereditary myopathies and dilated cardiomyopathy. Researchers have linked obscurin to titin due to their shared roles in maintaining sarcomeric structure. Mutations or dysregulations in either protein can lead to insufficient muscle function and structural cardiac issues implicating obscurin as a significant factor in these diseases.
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Homozygous: 1 bp insertion in exon 2.
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