Anti-MIRO2 antibody [EPR29118-10]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MIRO2 antibody [EPR29118-10] (AB321803)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling MIRO2 with ab321803 at 1/50 (10.16 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on human testis. The section was incubated with ab321803 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MIRO2 antibody [EPR29118-10] (AB321803)

Immunohistochemical analysis of paraffin-embedded Human endometrial carcinoma tissue labeling MIRO2 with ab321803 at 1/50 (10.16 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on human endometrial carcinoma. The section was incubated with ab321803 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MIRO2 antibody [EPR29118-10] (AB321803)

Immunohistochemical analysis of paraffin-embedded Human colon carcinoma tissue labeling MIRO2 with ab321803 at 1/50 (10.16 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on human colon carcinoma. The section was incubated with ab321803 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MIRO2 antibody [EPR29118-10] (AB321803)

Immunohistochemical analysis of paraffin-embedded Human endometrium tissue labeling MIRO2 with ab321803 at 1/50 (10.16 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on human endometrium. The section was incubated with ab321803 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MIRO2 antibody [EPR29118-10] (AB321803)

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling MIRO2 with ab321803 at 1/50 (10.16 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on mouse testis. The section was incubated with ab321803 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MIRO2 antibody [EPR29118-10] (AB321803)

Immunohistochemical analysis of paraffin-embedded Mouse Burkitt's lymophoma tissue labeling MIRO2 with ab321803 at 1/50 (10.16 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on mouse Burkitt's lymophoma. The section was incubated with ab321803 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MIRO2 antibody [EPR29118-10] (AB321803)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling MIRO2 with ab321803 at 1/50 (10.16 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on mouse colon. The section was incubated with ab321803 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

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Western blot - Anti-MIRO2 antibody [EPR29118-10] (AB321803)

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

Low expression : mouse lung (PMID : 15218247).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lane 1 : 10 seconds; Lanes 2-3 : 26 seconds; Lanes 4-6 : 15 seconds.

All lanes:

Western blot - Anti-MIRO2 antibody [EPR29118-10] (ab321803) 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 testis tissue lysate at 20 µg

Lane 5:

Mouse lung tissue lysate at 20 µg

Lane 6:

Mouse heart tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 80 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-MIRO2 antibody [EPR29118-10] (AB321803)

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

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-MIRO2 antibody [EPR29118-10] (ab321803) at 1/1000 dilution

Lane 1:

Human testis tissue lysate at 20 µg

Lane 2:

Human hippocampus tissue lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 80 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-MIRO2 antibody [EPR29118-10] (AB321803)

Blocking and diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of TBS.

The samples were run on a Bis-Tris gel under reducing conditions.

Western blot : Anti-MIRO2 antibody (ab) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta.

In Western blot, ab 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 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 [EPR29118-10] (ab321803) at 1/1000 dilution

Lane 1:

Wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

MIRO2 knockout HeLa whole cell lysate 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

Observed band size: 80 kDa,36 kDa

false