All lanes : Anti-Metabotropic Glutamate Receptor 3/MGLUR3 antibody [EPR24635-155] (ab281922) at 1/1000 dilution

Lane 1 : Mouse brain tissue lysate at 20 µg
Lane 2 : Mouse liver tissue lysate at 40 µg
Lane 3 : Rat brain tissue lysate at 20 µg
Lane 4 : Rat liver tissue lysate at 40 µg

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Predicted band size: 99 kDa
Observed band size: 100,250 kDa why is the actual band size different from the predicted?

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

Negative control: liver (PMID: 9001707).

The molecular weight observed is consistent with what has been described in the literature (PMID: 27130562).

Samples are non-boiled as boiling may cause protein aggregates.

Exposure time: 3.25 seconds.

 

Anti-Metabotropic Glutamate Receptor 3/MGLUR3 antibody [EPR24635-155] (ab281922) at 1/1000 dilution + Human brain tissue lysate at 20 µg

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

Predicted band size: 99 kDa
Observed band size: 100,250 kDa why is the actual band size different from the predicted?

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

The molecular weight observed is consistent with what has been described in the literature (PMID: 27130562).

Samples are non-boiled as boiling may cause protein aggregates.

Exposure time: 10 seconds.

 

All lanes : Anti-Metabotropic Glutamate Receptor 3/MGLUR3 antibody [EPR24635-155] (ab281922) at 1/1000 dilution

Lane 1 : HEK-293T (human embryonic kidney) transfected with an empty vector, containing a myc-His-tag®, whole cell lysate
Lane 2 : HEK-293T transfected with Metabotropic glutamate receptor 3 expression vector containing a myc-His-tag®, whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Predicted band size: 99 kDa
Observed band size: 100,250 kDa why is the actual band size different from the predicted?

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

The molecular weight observed is consistent with what has been described in the literature (PMID: 27130562).

Samples are non-boiled as boiling may cause protein aggregates.

Exposure time: 1 seconds

 

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labelling Metabotropic glutamate receptor 3/MGLUR3 with ab281922 at 1/1000 dilution (0.342 ug/ml) followed by a ready to use Rabbit specific IHC polymer Detection kit HRP/DAB (ab209101). Cytoplasmic staining on mouse cerebrum. The section was incubated with ab281922 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Immunohistochemical analysis of paraffin-embedded Mouse cerebellum tissue labelling Metabotropic glutamate receptor 3/MGLUR3 with ab281922 at 1/1000 dilution (0.342 ug/ml) followed by a ready to use Rabbit specific IHC polymer Detection kit HRP/DAB (ab209101). Cytoplasmic staining on mouse cerebellum. The section was incubated with ab281922 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labelling Metabotropic glutamate receptor 3/MGLUR3 with ab281922 at 1/1000 dilution (0.342 ug/ml) followed by a ready to use Rabbit specific IHC polymer Detection kit HRP/DAB (ab209101). Cytoplasmic staining on human cerebrum. The section was incubated with ab281922 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labelling Metabotropic glutamate receptor 3/MGLUR3 with ab281922 at 1/1000 dilution (0.342 ug/ml) followed by a ready to use Rabbit specific IHC polymer Detection kit HRP/DAB (ab209101). Cytoplasmic staining on rat cerebrum. The section was incubated with ab281922 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labelling Metabotropic glutamate receptor 3/MGLUR3 with ab281922 at 1/1000 dilution (0.342 ug/ml) followed by a ready to use Rabbit specific IHC polymer Detection kit HRP/DAB (ab209101). Negative control: No staining on mouse liver. The section was incubated with ab281922 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse hippocampus tissue labeling Metabotropic glutamate receptor 3/MGLUR3 with ab281922 at 1/100 dilution (3.42 ug/ml) followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). Cytoplasmic staining on mouse hippocampus is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488)at 1/1000 dilution.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat hippocampus tissue labeling Metabotropic glutamate receptor 3/MGLUR3 with ab281922 at 1/100 dilution (3.42 ug/ml) followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). Cytoplasmic staining on rat hippocampus is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488)at 1/1000 dilution.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse liver tissue labeling Metabotropic glutamate receptor 3/MGLUR3 with ab281922 at 1/100 dilution (6.84 ug/ml) followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). Negative control: No staining on mouse liver (PMID: 9001707) is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488)at 1/1000 dilution.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat liver tissue labeling Metabotropic glutamate receptor 3/MGLUR3 with ab281922 at 1/100 dilution (6.84 ug/ml) followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (Green). Negative control: No staining on rat liver (PMID: 9001707) is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488)at 1/1000 dilution.