Anti-Grp75/MOT antibody [EPR29618-542]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Grp75/MOT antibody [EPR29618-542] (AB322409)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling Grp75/MOT with ab322409 at 1/500 (1.028 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 NIH/3T3 cells (shown in green). 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 ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

-ve control 1 : ab322409 at a 1/500 dilution followed by ab150120 at a 1/1000 dilution.

-ve control 2 : anti-COX IV mouse monoclonal antibody - Mitochondrial Marker at a 1/200 dilution followed by ab150081 at a 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Grp75/MOT antibody [EPR29618-542] (AB322409)

Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling Grp75/MOT with ab322409 at 1/1000 (0.514 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse lung.
The section was incubated with ab322409 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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Grp75/MOT antibody [EPR29618-542] (AB322409)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling Grp75/MOT with ab322409 at 1/1000 (0.514 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse cerebrum.
The section was incubated with ab322409 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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Grp75/MOT antibody [EPR29618-542] (AB322409)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling Grp75/MOT with ab322409 at 1/1000 (0.514 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : No staining on mouse skeletal muscle
The section was incubated with ab322409 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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Grp75/MOT antibody [EPR29618-542] (AB322409)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Grp75/MOT with ab322409 at 1/1000 (0.514 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse kidney.
The section was incubated with ab322409 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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Grp75/MOT antibody [EPR29618-542] (AB322409)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Grp75/MOT with ab322409 at 1/1000 (0.514 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse liver.
The section was incubated with ab322409 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

• WB

Supplier Data

Western blot - Anti-Grp75/MOT antibody [EPR29618-542] (AB322409)

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

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

Exposure time : Lane 1 : 15 seconds; Lanes 2-3 : 114 seconds.

All lanes:

Western blot - Anti-Grp75/MOT antibody [EPR29618-542] (ab322409) at 1/2000 dilution

Lane 1:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 1 µg

Lane 2:

C2C12 (mouse myoblast) transfected with scrambled siRNA control whole cell lysate at 1 µg

Lane 3:

C2C12 transfected with siRNA specifically targeting Grp75/MOT whole cell lysate at 1 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 75 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-Grp75/MOT antibody [EPR29618-542] (AB322409)

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

Low expression tissue : skeletal muscle.

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

All lanes:

Western blot - Anti-Grp75/MOT antibody [EPR29618-542] (ab322409) at 1/1000 dilution

Lane 1:

Mouse kidney tissue lysate at 10 µg

Lane 2:

Mouse skeletal muscle tissue lysate at 10 µg

Lane 3:

Mouse liver tissue lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 75 kDa,36 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-Grp75/MOT antibody [EPR29618-542] (AB322409)

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

All lanes:

Western blot - Anti-Grp75/MOT antibody [EPR29618-542] (ab322409) at 1/1000 dilution

Lane 1:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 2:

RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 3:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 4:

MEF (mouse embryo fibroblast) whole cell lysate at 20 µg

Lane 5:

Mouse heart tissue lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 75 kDa

false

Exposure time: 8s