NF2 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 deletion in exon 1.
ACN, BANF, Bilateral acoustic neuroma, MERL_HUMAN, Merlin, Moesin ezrin radizin like, Moesin-ezrin-radixin-like protein, NF 2, Neurofibromatosis 2, Neurofibromatosis type 2, Neurofibromin-2, SCH, Schwannomerlin, Schwannomin
NF2 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 deletion 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 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.
The NF2 gene produces a protein known as Neurofibromin 2 often called Merlin. The NF2 protein or Merlin protein plays a mechanical role in the cell as a tumor suppressor by stabilizing cellular structure and regulating cell growth. Its molar mass is approximately 70 kDa. NF2 protein is found mainly in the nervous system particularly in Schwann cells which are important for nerve function.
The NF2 protein functions to control cell proliferation differentiation and motility. It associates with the cytoskeleton acting as a bridge between cell surface proteins and the actin cytoskeleton. Merlin as part of the ERM (ezrin radixin moesin) family contributes to the assembly of protein complexes at the cell membrane. Through its interactions it plays a significant role in maintaining proper cell signaling and structural integrity.
NF2 protein integrates into the Hippo signaling pathway which regulates organ size by controlling cell growth and apoptosis. Within this pathway Merlin interacts with members such as YAP1 and LATS1/2 moderating their activity to prevent over-proliferation of cells. NF2 also connects with the mTOR pathway which is critical for cell metabolism and growth further emphasizing its regulatory functions in cellular homeostasis.
Mutations or deletions in the NF2 gene are associated with neurofibromatosis type 2 a condition that causes benign tumors on nerve tissues known as schwannomas. NF2 protein is linked to other proteins such as angiogenesis factors contributing to the development of these tumors. Schwannomatosis another disorder related to NF2 involves nerve sheath tumor formation with Merlin mutation playing a role indicating its importance in nervous system health.
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All lanes: Anti-NF2 / Merlin antibody [AF1G4] (ab88957) at 1/2000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: NF2 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human NF2 (Merlin) knockout HeLa cell line (ab261796)
Performed under reducing conditions.
Anti-NF2 / Merlin antibody [EPR2573(2)] ab109244 was shown to react with NF2 in wild-type HeLa cells in western blot with loss of signal observed in NF2 knockout cell line ab261796 (NF2 knockout cell lysate Human NF2 (Merlin) knockout HeLa cell lysate ab257179). Wild-type and NF2 knockout HeLa cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with Anti-NF2 / Merlin antibody [EPR2573(2)] ab109244 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4° at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (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-NF2 / Merlin antibody [EPR2573(2)] (Anti-NF2 / Merlin antibody [EPR2573(2)] ab109244) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: NF2 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human NF2 (Merlin) knockout HeLa cell line (ab261796)
Performed under reducing conditions.
Predicted band size: 70 kDa
Observed band size: 60 kDa
Homozygous: 1 bp deletion in exon 1.
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