Human MFN2 (Mitofusin 2) knockout HEK-293 cell line
- Advanced Validation
- What is this?
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(1 Publication)
MFN2 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 10 bp deletion.
View Alternative Names
CMT2A, CMT2A2, CPRP 1, EC 3.6.5.-, Fzo, HSG, Hypertension related protein 1, KIAA0214, MARF, MFN2_HUMAN, Mitochondrial assembly regulatory factor, Mitofusin-2, Transmembrane GTPase MFN2, hyperplasia suppressor gene
- WB
Lab
Western blot - Human MFN2 (Mitofusin 2) knockout HEK-293 cell line (AB260861)
Western blot : Anti-MFN2 antibody [EPR19796] (ab205236) staining at 1/2000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab205236 was shown to bind specifically to MFN2. A band was observed at 80 kDa in wild-type HEK-293 cell lysates with no signal observed at this size in MFN2 knockout cell line. To generate this image, wild-type and MFN2 knockout HEK-293 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes:
Western blot - Anti-Mitofusin 2 antibody [EPR19796] (<a href='/en-us/products/primary-antibodies/mitofusin-2-antibody-epr19796-ab205236'>ab205236</a>) at 1/2000 dilution
Lane 1:
Wild-type HEK-293 cell lysate at 20 µg
Lane 2:
MFN2 knockout HEK-293 cell lysate at 20 µg
Lane 3:
HeLa cell lysate at 20 µg
Lane 4:
PC-3 cell lysate at 20 µg
Secondary
All lanes:
Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Observed band size: 80 kDa
false
- WB
Lab
Western blot - Human MFN2 (Mitofusin 2) knockout HEK-293 cell line (AB260861)
Western blot : Anti-MFN2 antibody [NIAR164] (ab124773) staining at 1/5000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab124773 was shown to bind specifically to MFN2. A band was observed at 64 kDa in wild-type HEK-293 cell lysates with no signal observed at this size in MFN2 knockout cell line. To generate this image, wild-type and MFN2 knockout HEK-293 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes:
Western blot - Anti-Mitofusin 2 antibody [NIAR164] (<a href='/en-us/products/primary-antibodies/mitofusin-2-antibody-niar164-ab124773'>ab124773</a>) at 1/5000 dilution
Lane 1:
Wild-type HEK-293 cell lysate at 20 µg
Lane 2:
Western blot - Human MFN2 (Mitofusin 2) knockout HEK-293 cell line (ab260861)
Lane 2:
MFN2 knockout HEK-293 cell lysate at 20 µg
Lane 3:
HeLa cell lysate at 20 µg
Lane 4:
PC-3 cell lysate at 20 µg
Secondary
Lanes 1 - 4:
Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution
Lanes 1 - 4:
Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Observed band size: 64 kDa
false
- WB
Lab
Western blot - Human MFN2 (Mitofusin 2) knockout HEK-293 cell line (AB260861)
Lanes 1 - 4 : Merged signal (red and green). Green - ab205236 observed at 86 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab205236 was shown to recognize MFN2 (Mitofusin 2) in wild-type HEK-293 cells as signal was lost at the expected MW in MFN2 knockout cell line ab260861 (knockout cell lysate ab261653). Additional cross-reactive bands were observed in the wild-type and knockout samples. Wild-type and MFN2 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% milk. ab205236 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/2000 dilution and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-Mitofusin 2 antibody [EPR19796] (<a href='/en-us/products/primary-antibodies/mitofusin-2-antibody-epr19796-ab205236'>ab205236</a>) at 1/2000 dilution
Lane 1:
Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
Lane 2:
MFN2 knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
Lane 2:
Western blot - Human MFN2 (Mitofusin 2) knockout HEK-293 cell line (ab260861)
Lane 3:
Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
Lane 4:
HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Predicted band size: 86 kDa
false
- NGS
Supplier Data
Next Generation Sequencing - Human MFN2 (Mitofusin 2) knockout HEK-293 cell line (AB260861)
10 bp deletion after Ile112 of the WT protein
Reactivity data
Product details
Recommended control: Human wild-type HEK-293 cell line (ab259776). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
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
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
Mitofusin 2 ensures the proper distribution of mitochondria within cells and regulates mitochondrial metabolism. It is a critical component of the mitochondrial fusion machinery and works closely with its homolog Mitofusin 1 (MFN1). Together they form a complex that facilitates the physical merging of mitochondrial membranes. This process is essential for mitochondrial dynamics which include not only fusion but also fission and biogenesis.
Pathways
The protein part of the fusion machinery integrates into multiple essential biological pathways including energy metabolism and apoptosis regulation. It participates in the mitochondrial fusion pathway and the PGC-1α pathway for mitochondrial biogenesis. Mitofusin 2 interacts with proteins such as PINK1 and Parkin that are known to play roles in mitophagy a process that targets damaged mitochondria for degradation indicating its involvement in maintaining mitochondrial quality control.
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
Publications (1)
Recent publications for all applications. Explore the full list and refine your search
Frontiers in aging neuroscience 14:990679 PubMed36337694
2022
Applications
Unspecified application
Species
Unspecified reactive species
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Product promise
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