人RHOT2 (MIRO2) knockout HeLa cell line
Human RHOT2 (MIRO2) knockout HeLa cell line
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RHOT2 KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 2 and 2 bp deletion in exon 2.
查看别名
ARHT2, C16orf39, MIRO2_HUMAN, Mitochondrial Rho GTPase 2, RASL, RHOT2, Ras homolog gene family member T2, hMiro-2
- Sanger seq
Unknown
Sanger Sequencing - Human RHOT2 (MIRO2) knockout HeLa cell line (AB265801)
Allele-2 : 1 bp deletion in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human RHOT2 (MIRO2) knockout HeLa cell line (AB265801)
Allele-1 : 2 bp deletion in exon 2.
- ICC/IF
Supplier Data
Immunocytochemistry/ Immunofluorescence - Human RHOT2 (MIRO2) knockout HeLa cell line (AB265801)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RHOT2 KO HeLa (RHOT2 knockout human cervical adenocarcinoma epithelial cell), ab265801 cells labelling MIRO2 with ab322962 at 1/500 (0.906 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).
Confocal image showing mitochondrial staining in wildtype HeLa cells (shown in green), showing no staining in RHOT2 knockout HeLa cells. The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain tubulin at 1/200 2.5 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
-ve control 1 : ab322962 at 1/500 dilution, followed byab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution. -ve control 2 : anti-COX IV mouse monoclonal - Mitochondrial Marker antibody at 1/200 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.
- ICC/IF
Supplier Data
Immunocytochemistry/ Immunofluorescence - Human RHOT2 (MIRO2) knockout HeLa cell line (AB265801)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RHOT2 KO HeLa (RHOT2 knockout human cervical adenocarcinoma epithelial cell), ab265801 cells labelling MIRO2 with ab320739 at 1/50 (9.12 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing mitochondrial staining in wildtype HeLa cells (shown in green), showing no staining in RHOT2 knockout HeLa cells. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 (2 ug/ml) dilution (Magenta). The Nuclear counterstain was DAPI (Blue). -ve control 1 : ab320739 at 1/50 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 (2 ug/ml) dilution. -ve control 2 : anti-COX IV mouse monoclonal antibody at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.
产品详情
Recommended control: Human wild-type HeLa cell line (ab255928). 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.
规格
性能和储存信息
基因名称
基因编辑类型
基因编辑方法
敲除验证
运输条件
推荐的短期储存条件
推荐的长期储存条件
处理步骤
初始处理指南
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.
传代培养指南
- 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.
培养基
DMEM (High Glucose) + 10% FBS
低温储藏试剂
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
补充信息
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
MIRO2 contributes to the maintenance of mitochondrial network and quality within cells. It serves as a component of the calcium-sensing mitochondrial transport machinery and interacts with the transport protein complex including adaptor proteins like Milton and the motor protein kinesin. Through these interactions MIRO2 helps in coupling mitochondrial dynamics to cellular events by facilitating calcium-dependent docking and release of mitochondria. This function indicates the importance of MIRO2 in energy-demanding cellular processes.
Pathways
MIRO2 integrates into the mitochondrial transport and positioning pathways. It associates with the PINK1/Parkin pathway which is essential for mitochondrial quality control and turnover. The pathway leads to the balance of mitochondrial fission and fusion assisting in the removal of damaged mitochondria. Moreover MIRO2 collaborates with proteins like Mitofusins in these pathways to ensure efficient mitochondrial dynamics and bioenergetics.
质量控制
STR 分析
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
细胞培养
生物安全等级
EU: 2 US: 2
贴壁/悬浮
Adherent
性别
Female
Complementary Products
Primary Antibodies
AB321803
Anti-MIRO2 antibody [EPR29118-10]
BOND RX™ Validated|20ul selling size|Recombinant|RabMAb|KO Validated
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MIRO2 antibody [EPR29118-10] (AB321803)](https://content.abcam.com/products/images/miro2-antibody-epr29118-10-ab321803--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img441178.jpg)
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