RHOT2 KO cell lysate available now. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon2 and 2 bp deletion in exon2.
Images
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon2 and 2 bp deletion in exon2.
Alternative names
ARHT2, C16orf39, MIRO2_HUMAN, Mitochondrial Rho GTPase 2, RASL, RHOT2, Ras homolog gene family member T2, hMiro-2
What's included?
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RHOT2 KO cell lysate available now. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon2 and 2 bp deletion in exon2.
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon2 and 2 bp deletion in exon2.
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- RHOT2
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 2 US: 2
- Adherent/suspension
- Adherent
- Gender
- Female
Storage
- Shipped at conditions
- Ambient - Can Ship with Ice
- Appropriate short-term storage conditions
- -20°C
- Appropriate long-term storage conditions
- -20°C
Notes
Knockout cell lysate achieved by CRISPR/Cas9.
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
MIRO2 also known as Rhot2 or Mitochondrial Rho GTPase 2 is a mitochondrial Rho GTPase with a molecular weight of approximately 69 kDa. This protein belongs to the Rho family of GTPases and exists mainly on the mitochondrial outer membrane. It acts as a critical regulator of mitochondrial trafficking and dynamics. MIRO2 plays a pivotal role in the regulation of mitochondrial transport along the cytoskeleton by interacting with adaptor proteins. It features two EF-hand motifs and two GTPase domains that are essential for its function.
- 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.
- Associated diseases and disorders
MIRO2 has links to neurodegenerative diseases like Parkinson's disease. Dysfunction in the PINK1/Parkin pathway where MIRO2 participates leads to impaired mitophagy and neuronal cell death. Additionally imbalances in MIRO2 activity are implicated in the progression of certain types of cancer. In these contexts MIRO2 interacts with related proteins such as PINK1 and Mitofusins which play roles in disease mechanisms that involve mitochondrial dysfunction or aberrant cellular metabolism.
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4 product images
Sanger Sequencing - Human RHOT2 (MIRO2) knockout HeLa cell lysate (ab257639)
Allele-2: 1 bp deletion in exon2
Sanger Sequencing - Human RHOT2 (MIRO2) knockout HeLa cell lysate (ab257639)
Allele-1: 2 bp deletion in exon2
Western blot - Human RHOT2 (MIRO2) knockout HeLa cell lysate (ab257639)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The samples were run on a Bis-Tris gel under reducing conditions.
Western blot: Anti-MIRO2 antibody (Anti-MIRO2 antibody [RM2077] ab322962) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody 6C5 loading control staining at 1/20000 dilution, shown in magenta.
In Western blot, Anti-MIRO2 antibody [RM2077] ab322962 was shown to bind specifically to MIRO2. Target of interest was observed at 80 kDa in wild-type HeLa cell lysates (lane 1) with no signal observed at this size in MIRO2 knockout cell line (lane 2, knockout cell line Human RHOT2 (MIRO2) knockout HeLa cell line ab265801 / knockout cell lysate ab257639). To generate this image, samples were first run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in a fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed in TBS-T, incubated with secondary antibodies Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution for 1 h at room temperature, washed again then imaged.
All lanes: Western blot - Anti-MIRO2 antibody [RM2077] (Anti-MIRO2 antibody [RM2077] ab322962) at 1/1000 dilution
Lane 1: Wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2: Western blot - Human RHOT2 (MIRO2) knockout HeLa cell lysate (ab257639) at 20 µg
Lane 3: 293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg
Lane 4: K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg
SecondaryAll lanes: Goat Anti-Rabbit IgG H&L (800CW) and Goat Anti-Mouse IgG H&L (680RD) at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 80 kDa
Western blot - Human RHOT2 (MIRO2) knockout HeLa cell lysate (ab257639)
Blocking buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of TBS.
Diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBST
The samples were run on a Bis-Tris gel under reducing conditions.
Western blot: Anti-MIRO2 antibody (Anti-MIRO2 antibody [EPR29118-85] ab320739) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in magenta.
In Western blot, Anti-MIRO2 antibody [EPR29118-85] ab320739 was shown to bind specifically to MIRO2. Target of interest was observed at 80 kDa in wild-type HeLa cell lysates (lane 1) with no signal observed at this size MIRO2 knockout cell line (lane 2, knockout cell line Human RHOT2 (MIRO2) knockout HeLa cell line ab265801 / knockout cell lysate ab257639). To generate this image, samples were first run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in a fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed in TBS-T, incubated with secondary antibodies Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution for 1 h at room temperature, washed again then imaged.
Exposure time: N/AAll lanes: Western blot - Anti-MIRO2 antibody [EPR29118-85] (Anti-MIRO2 antibody [EPR29118-85] ab320739) at 1/1000 dilution
Lane 1: Wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2: Western blot - Human RHOT2 (MIRO2) knockout HeLa cell lysate (ab257639) at 20 µg
Lane 3: 293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg
Lane 4: K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg
SecondaryAll lanes: Goat Anti-Rabbit IgG H&L (800CW) and Goat Anti-Mouse IgG H&L (680RD) at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 80 kDa
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