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Knockout Tested Rabbit Recombinant Monoclonal MIRO2 antibody. Carrier free. Suitable for WB, ICC/IF and reacts with Recombinant fragment - Human, Human samples.

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Images

Western blot - Anti-MIRO2 antibody [EPR29118-85] - BSA and Azide free (AB320740), expandable thumbnail
  • Western blot - Anti-MIRO2 antibody [EPR29118-85] - BSA and Azide free (AB320740), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-MIRO2 antibody [EPR29118-85] - BSA and Azide free (AB320740), expandable thumbnail
  • Western blot - Anti-MIRO2 antibody [EPR29118-85] - BSA and Azide free (AB320740), expandable thumbnail

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Constituents: 100% PBS

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBICC/IF
Human
Tested
Tested
Mouse
Not recommended
Not recommended
Recombinant fragment - Human
Tested
Not recommended

Tested
Tested

Species

Recombinant fragment - Human, Human

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Mouse

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Mouse, Recombinant fragment - Human

Dilution info

-

Notes

-

Target data

Function

Mitochondrial GTPase involved in mitochondrial trafficking (PubMed:16630562, PubMed:22396657). Probably involved in control of anterograde transport of mitochondria and their subcellular distribution (PubMed:22396657).

Alternative names

Mitochondrial Rho GTPase 2, MIRO-2, hMiro-2, Ras homolog gene family member T2, C16orf39, ARHT2, RHOT2

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Knockout Tested Rabbit Recombinant Monoclonal MIRO2 antibody. Carrier free. Suitable for WB, ICC/IF and reacts with Recombinant fragment - Human, Human samples.

Alternative names

Mitochondrial Rho GTPase 2, MIRO-2, hMiro-2, Ras homolog gene family member T2, C16orf39, ARHT2, RHOT2

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free

Yes

Clone number

EPR29118-85

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate long-term storage conditions

+4°C

Notes

ab320740 is the carrier-free version of Anti-MIRO2 antibody [EPR29118-85] ab320739.

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

Supplementary info

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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

4 product images

  • Western blot - Anti-MIRO2 antibody [EPR29118-85] - BSA and Azide free (ab320740), expandable thumbnail

    Western blot - Anti-MIRO2 antibody [EPR29118-85] - BSA and Azide free (ab320740)

    This data was developed using Anti-MIRO2 antibody [EPR29118-85] ab320739, the same antibody clone in a different buffer formulation.

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    Lane 3 are applied with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 and lanes 1-2 are applied with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000.

    Lanes 1-2 of this blot were developed using a high sensitivity ECL substrate.

    The high-sensitivity ECL substrate allows for the detection of proteins in the mid-femtogram range.

    Exposure time: Lanes 1-2: 180 seconds; Lane 3: 6 seconds.

    All lanes: Western blot - Anti-MIRO2 antibody [EPR29118-85] (Anti-MIRO2 antibody [EPR29118-85] ab320739) at 1/1000 dilution

    Lane 1: Human testis tissue lysate at 20 µg

    Lane 2: Human hippocampus tissue lysate at 20 µg

    Lane 3: HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg

    Secondary

    Lanes 1 - 2: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

    Lane 3: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Observed band size: 80 kDa

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    Lane 3 are applied with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 and lanes 1-2 are applied with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000.

    Lanes 1-2 of this blot were developed using a high sensitivity ECL substrate.

    The high-sensitivity ECL substrate allows for the detection of proteins in the mid-femtogram range.

    Exposure time: Lanes 1-2: 180 seconds; Lane 3: 6 seconds.

  • Western blot - Anti-MIRO2 antibody [EPR29118-85] - BSA and Azide free (ab320740), expandable thumbnail

    Western blot - Anti-MIRO2 antibody [EPR29118-85] - BSA and Azide free (ab320740)

    This data was developed using Anti-MIRO2 antibody [EPR29118-85] ab320739, the same antibody clone in a different buffer formulation.

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    This antibody does not cross-react with recombinant human MIRO1 by western blot.

    In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade  (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204) staining at 1/5000 dilution.

    All lanes: Western blot - Anti-MIRO2 antibody [EPR29118-85] (Anti-MIRO2 antibody [EPR29118-85] ab320739) at 1/1000 dilution

    Lane 1: His-tagged human MIRO2 fragment at 10 ng

    Lane 2: His-tagged human MIRO1 fragment at 10 ng

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Observed band size: 20 kDa, 21 kDa

    Exposure time: 10s

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    This antibody does not cross-react with recombinant human MIRO1 by western blot.

    In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade  (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204) staining at 1/5000 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-MIRO2 antibody [EPR29118-85] - BSA and Azide free (ab320740), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-MIRO2 antibody [EPR29118-85] - BSA and Azide free (ab320740)

    This data was developed using Anti-MIRO2 antibody [EPR29118-85] ab320739, the same antibody clone in a different buffer formulation.

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RHOT2 KO HeLa (RHOT2 knockout human cervical adenocarcinoma epithelial cell), Human RHOT2 (MIRO2) knockout HeLa cell line ab265801 cells labelling MIRO2 with Anti-MIRO2 antibody [EPR29118-85] ab320739 at 1/50 (9.12 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed 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: Anti-MIRO2 antibody [EPR29118-85] ab320739 at 1/50 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

  • Western blot - Anti-MIRO2 antibody [EPR29118-85] - BSA and Azide free (ab320740), expandable thumbnail

    Western blot - Anti-MIRO2 antibody [EPR29118-85] - BSA and Azide free (ab320740)

    This data was developed using Anti-MIRO2 antibody [EPR29118-85] ab320739, the same antibody clone in a different buffer formulation.

    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 Human RHOT2 (MIRO2) knockout HeLa 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/A

    All 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 (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

    Secondary

    All 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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Product protocols

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