Anti-GPR56 antibody [RM2094]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-GPR56 antibody [RM2094] (AB326042)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized B16-F0 (mouse melanoma epithelial-like cell) F9 (mouse embryonal carcinoma epithelial cell) cells labelling GPR56 with ab326042 at 1/50 (9.98 µg/ml) dilution , followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in B16-F0 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Low expression : F9. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab326042 at 1/50 dilution, followed by ab150120 at 1/1000 dilution.-ve control 2 : ab7291 at 1/1000 dilution, followed by ab150081 at 1/1000 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-GPR56 antibody [RM2094] (AB326042)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized B16-F10 (mouse skin melanoma cell) Neuro-2a (mouse neuroblastoma neuroblast) cells labelling GPR56 with ab326042 at 1/50 (9.98 µg/ml) dilution , followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in B16-F10 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Low expression : Neuro-2a. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab326042 at 1/50 dilution, followed by ab150120 at 1/1000 dilution.-ve control 2 : ab7291 at 1/1000 dilution, followed by ab150081 at 1/1000 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GPR56 antibody [RM2094] (AB326042)

Immunohistochemical analysis of paraffin-embedded Mouse E14.5 (Brain section) tissue labeling GPR56 with ab326042 at 1/100 (4.99 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse E14.5 brain. (A) Low-powered (magnification, x20) and (B) high-powered (magnification, x400) microscopic images. (PMID : 21768377).

The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GPR56 antibody [RM2094] (AB326042)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling GPR56 with ab326042 at 1/100 (4.99 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse kidney (PMID : 17932623).

The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GPR56 antibody [RM2094] (AB326042)

Immunohistochemical analysis of paraffin-embedded Mouse brain (Coronal section) tissue labeling GPR56 with ab326042 at 1/100 (4.99 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse brain. (A) Low-powered (magnification, x20) and (B) high-powered (magnification, x400) microscopic images. (PMID : 17932623).

The primary antibody was incubated 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

• Flow Cyt

Lab

Flow Cytometry - Anti-GPR56 antibody [RM2094] (AB326042)

Flow cytometric analysis of Neuro-2a (mouse neuroblastoma neuroblast, Left) B16-F10 (mouse skin melanoma cell, Right) cells labelling GPR56 with ab326042 at 1/500 dilution (0.1ug) / Red compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells. Low expression : Neuro-2a.

• Flow Cyt

Lab

Flow Cytometry - Anti-GPR56 antibody [RM2094] (AB326042)

Flow cytometric analysis of F9 (mouse embryonal carcinoma epithelial cell, Left) B16-F0 (mouse melanoma epithelial-like cell, Right) cells labelling GPR56 with ab326042 at 1/500 dilution (0.1ug) / Red compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells. Low expression : F9.

• WB

Lab

Western blot - Anti-GPR56 antibody [RM2094] (AB326042)

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

Low expression : Neuro-2a, F9.

This antibody could detect 2 isoforms.

The identity of the higher MW band at approximately 250 kDa (in lane 3) is unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-GPR56 antibody [RM2094] (ab326042) at 1/1000 dilution

Lane 1:

B16-F10 (mouse skin melanoma cell) whole cell lysate at 20 µg

Lane 2:

B16-F0 (mouse melanoma epithelial-like cell) whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate at 20 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 1/20000 dilution

Observed band size: 57 kDa,75 kDa,36 kDa

false

Exposure time: 136s

• WB

Lab

Western blot - Anti-GPR56 antibody [RM2094] (AB326042)

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

Low expression : liver, skeletal muscle, heart, lung.

This antibody could detect 2 isoforms.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-GPR56 antibody [RM2094] (ab326042) at 1/1000 dilution

Lane 1:

Mouse E14 brain tissue lysate at 20 µg

Lane 2:

Mouse brain tissue lysate at 20 µg

Lane 3:

Mouse kidney tissue lysate at 20 µg

Lane 4:

Mouse liver tissue lysate at 20 µg

Lane 5:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 6:

Mouse heart tissue lysate at 20 µg

Lane 7:

Mouse lung tissue lysate at 20 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 1/20000 dilution

Observed band size: 57 kDa,75 kDa,36 kDa

false

Exposure time: 136s

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GPR56 antibody [RM2094] (AB326042)

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling GPR56 with ab326042 at 1/100 (4.99 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : No staining on mouse cardiac muscle (PMID : 17932623).

The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GPR56 antibody [RM2094] (AB326042)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling GPR56 with ab326042 at 1/100 (4.99 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : No staining on mouse skeletal muscle.

The primary antibody was incubated 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