Rabbit Recombinant Multiclonal MIRO2 antibody. Suitable for WB, IHC-P, ICC/IF and reacts with Recombinant fragment - Human, Human, Mouse, Rat samples.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Multiclonal
WB | IHC-P | ICC/IF | Flow Cyt (Intra) | IP | |
---|---|---|---|---|---|
Human | Tested | Tested | Tested | Not recommended | Not recommended |
Mouse | Tested | Not recommended | Not recommended | Not recommended | Not recommended |
Rat | Tested | Not recommended | Expected | Not recommended | Not recommended |
Recombinant fragment - Human | Tested | Not recommended | Not recommended | Not recommended | Not recommended |
Species | Dilution info | Notes |
---|---|---|
Species Recombinant fragment - Human | Dilution info 1/1000 | Notes - |
Species Human | Dilution info 1/1000 | Notes - |
Species Mouse | Dilution info 1/1000 | Notes - |
Species Rat | Dilution info 1/1000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/100 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat, Recombinant fragment - Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/500 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Recombinant fragment - Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human, Mouse, Rat, Recombinant fragment - Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human, Mouse, Rat, Recombinant fragment - Human | Dilution info - | Notes - |
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).
ARHT2, C16orf39, C16orf39, ARHT2, RHOT2, Mitochondrial Rho GTPase 2, MIRO-2, hMiro-2, Ras homolog gene family member T2
Rabbit Recombinant Multiclonal MIRO2 antibody. Suitable for WB, IHC-P, ICC/IF and reacts with Recombinant fragment - Human, Human, Mouse, Rat samples.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Multiclonal
RM2077
Affinity purification Protein A
Unsuitable for mouse ICC.
Unsuitable for mouse and rat IHC-P
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
This product is a recombinant multiclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our 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.
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 ab322962 at 1/500 (0.906 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. 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 antibody - Mitochondrial Marker at 1/200 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 dilution.
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 [RM2077] (ab322962) at 1/1000 dilution
Lane 1: His-tagged human MIRO2 fragment at 10 ng
Lane 2: His-tagged human MIRO1 fragment at 10 ng
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: 21 kDa, 23 kDa
Exposure time: 15s
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Low expression: mouse lung (PMID: 15218247).
The identity of the band lower than 75 kDa is unknown.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
Exposure time: Lane 1: 10 seconds; Lanes 2-3: 81 seconds; Lanes 4-6: 37 seconds.
All lanes: Western blot - Anti-MIRO2 antibody [RM2077] (ab322962) at 1/1000 dilution
Lane 1: HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg
Lane 3: MEF (mouse embryo fibroblast) whole cell lysate at 20 µg
Lane 4: Mouse heart tissue lysate at 20 µg
Lane 5: Mouse lung tissue lysate at 20 µg
Lane 6: Mouse testis tissue lysate at 20 µg
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: 80 kDa, 36 kDa
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The identity of the band lower than 75 kDa is unknown.
Exposure time: Lane 1: 81 seconds; Lane 2: 180 seconds.
All lanes: Western blot - Anti-MIRO2 antibody [RM2077] (ab322962) at 1/1000 dilution
Lane 1: Human hippocampus tissue lysate at 20 µg
Lane 2: Human testis tissue lysate at 20 µg
All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Observed band size: 80 kDa
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 (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, 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 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.
All lanes: Western blot - 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 (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
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
Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling MIRO2 with ab322962 at 1/100 (4.53 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on human colon. The section was incubated with ab322962 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling MIRO2 with ab322962 at 1/100 (4.53 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on human breast carcinoma. The section was incubated with ab322962 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemical analysis of paraffin-embedded Human colon carcinoma tissue labeling MIRO2 with ab322962 at 1/100 (4.53 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on human colon carcinoma. The section was incubated with ab322962 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
All lanes: Western blot - Anti-MIRO2 antibody [RM2077] (ab322962) at 1/1000 dilution
Lane 1: PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg
Lane 2: Rat testis tissue lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Exposure time: 37s
Immunohistochemical analysis of paraffin-embedded Human pancreatic carcinoma tissue labeling MIRO2 with ab322962 at 1/100 (4.53 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on human pancreatic carcinoma. The section was incubated with ab322962 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemical analysis of paraffin-embedded Human endometrium tissue labeling MIRO2 with ab322962 at 1/100 (4.53 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on human endometrium. The section was incubated with ab322962 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
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