Human NF2 (Merlin) knockout HeLa cell line
- Advanced Validation
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NF2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 1. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
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
- WB
Lab
Western blot - Human NF2 (Merlin) knockout HeLa cell line (AB261796)
Lanes 1 - 2 : Merged signal (red and green). Green - ab109244 observed at 60 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
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 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 ab109244 and 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (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)] (<a href='/en-us/products/primary-antibodies/nf2-merlin-antibody-epr25732-ab109244'>ab109244</a>) 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)
Predicted band size: 70 kDa
Observed band size: 60 kDa
false
- WB
Lab
Western blot - Human NF2 (Merlin) knockout HeLa cell line (AB261796)
Lanes 1 - 2 : Merged signal (red and green). Green - ab88957 observed at 60 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37kDa.
ab88957 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 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 ab88957 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4°C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively.. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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)
false
- Sanger seq
Unknown
Sanger Sequencing - Human NF2 (Merlin) knockout HeLa cell line (AB261796)
Homozygous : 1 bp deletion in exon 1.
Reactivity data
Product details
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
Initial handling guidelines
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.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
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.
Pathways
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
Primary Antibodies
AB109244
Anti-NF2 / Merlin antibody [EPR2573(2)]
RabMAb|Recombinant|KO Validated
![Western blot - Anti-NF2 / Merlin antibody [EPR2573(2)] (AB109244)](https://content.abcam.com/products/images/nf2-merlin-antibody-epr25732-ab109244--western-blot-img165311.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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