Anti-Cannabinoid Receptor I antibody [EPR30843-34]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cannabinoid Receptor I antibody [EPR30843-34] (AB325797)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling Cannabinoid Receptor I with ab325797 at 1/2000 (0.241 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human cerebrum.

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-Cannabinoid Receptor I antibody [EPR30843-34] (AB325797)

Immunohistochemical analysis of paraffin-embedded Human hippocampus tissue labeling Cannabinoid Receptor I with ab325797 at 1/2000 (0.241 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human hippocampus.

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-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-Cannabinoid Receptor I antibody [EPR30843-34] (AB325797)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat cerebellum (fresh frozen) tissue labeling Cannabinoid Receptor I with ab325797 at 1/200 (2.41 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 µg/mL dilution (Green).

Panel A : merged staining of anti-CNR1 (ab325797, green), anti-NeuN (ab190565, magenta) on rat cerebellum. Panel B : anti-CNR1 stained on rat cerebellum. Panel C : anti-NeuN stained in neurons of rat cerebellum. The section was incubated in two rounds of staining : in the order of ab325797 and ab190565, for 1 hr at room temperature. The nuclear counterstain was DAPI (Blue). The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cannabinoid Receptor I antibody [EPR30843-34] (AB325797)

Immunohistochemical analysis of paraffin-embedded Mouse cerebellum tissue labeling Cannabinoid Receptor I with ab325797 at 1/2000 (0.241 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse cerebellum.

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-Cannabinoid Receptor I antibody [EPR30843-34] (AB325797)

Immunohistochemical analysis of paraffin-embedded Mouse hippocampus tissue labeling Cannabinoid Receptor I with ab325797 at 1/2000 (0.241 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse hippocampus (PMID : 40999232).

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-Cannabinoid Receptor I antibody [EPR30843-34] (AB325797)

Immunohistochemical analysis of paraffin-embedded Rat cerebellum tissue labeling Cannabinoid Receptor I with ab325797 at 1/2000 (0.241 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat cerebellum.

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-Cannabinoid Receptor I antibody [EPR30843-34] (AB325797)

Immunohistochemical analysis of paraffin-embedded Rat hippocampus tissue labeling Cannabinoid Receptor I with ab325797 at 1/2000 (0.241 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat hippocampus (PMID : 26578900).

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-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-Cannabinoid Receptor I antibody [EPR30843-34] (AB325797)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebellum (fresh frozen) tissue labeling Cannabinoid Receptor I with ab325797 at 1/200 (2.41 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 µg/mL dilution (Green).

Panel A : merged staining of anti-CNR1 (ab325797, green), anti-NeuN (ab190565, magenta) on mouse cerebellum. Panel B : anti-CNR1 stained on mouse cerebellum. Panel C : anti-NeuN stained in neurons of mouse cerebellum. The section was incubated in two rounds of staining : in the order of ab325797 and ab190565, for 1 hr at room temperature. The nuclear counterstain was DAPI (Blue). The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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

• WB

Lab

Western blot - Anti-Cannabinoid Receptor I antibody [EPR30843-34] (AB325797)

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

Low expression : kidney, liver(PMID : 25760323).

The samples are unboiled.

The band pattern observed is consistent with what has been described in the literature (PMID : 34833992).

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

All lanes:

Western blot - Anti-Cannabinoid Receptor I antibody [EPR30843-34] (ab325797) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 40 µg

Lane 2:

Human kidney tissue lysate at 40 µg

Lane 3:

Mouse brain tissue lysate at 40 µg

Lane 4:

Mouse cerebellum tissue lysate at 40 µg

Lane 5:

Mouse kidney tissue lysate at 40 µg

Lane 6:

Mouse liver tissue lysate at 40 µg

Lane 7:

Rat brain tissue lysate at 40 µg

Lane 8:

Rat cerebellum tissue lysate at 40 µg

Lane 9:

Rat kidney tissue lysate at 40 µg

Secondary

All lanes:

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

Observed band size: 37 kDa,50 kDa,36 kDa

false

Exposure time: 6s

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cannabinoid Receptor I antibody [EPR30843-34] (AB325797)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling Cannabinoid Receptor I with ab325797 at 1/2000 (0.241 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : no staining on rat kidney.

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-Cannabinoid Receptor I antibody [EPR30843-34] (AB325797)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human embryonic kidney epithelial cell) transfected with a CNR1 expression vector containing a his tag (B) HEK-293T transfected with a CNR2 expression vector containing a his tag (C) HEK-293T cells transfected with empty vector containing a myc-His-tag tissue labeling Cannabinoid Receptor I with ab325797 at 1/8000 (0.06 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining (A) HEK-293T transfected with a CNR1 expression vector containing a his tag, no staining on (B) HEK-293T transfected with a CNR2 expression vector containing a his tag and (C) HEK-293T cells transfected with empty vector containing a myc-His-tag.

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-Cannabinoid Receptor I antibody [EPR30843-34] (AB325797)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Cannabinoid Receptor I with ab325797 at 1/2000 (0.241 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : No staining on human kidney.

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-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-Cannabinoid Receptor I antibody [EPR30843-34] (AB325797)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse kidney (fresh frozen) tissue labeling Cannabinoid Receptor I with ab325797 at 1/200 (2.41 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 µg/mL dilution (Green).

Low expression : confocal image showing no staining on mouse kidney (PMID : 25760323). The nuclear counterstain was DAPI (Blue). The section was incubated with ab325797 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-Cannabinoid Receptor I antibody [EPR30843-34] (AB325797)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat kidney (fresh frozen) tissue labeling Cannabinoid Receptor I with ab325797 at 1/200 (2.41 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 µg/mL dilution (Green).

Low expression : confocal image showing no staining on rat kidney. The nuclear counterstain was DAPI (Blue). The section was incubated with ab325797 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cannabinoid Receptor I antibody [EPR30843-34] (AB325797)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Cannabinoid Receptor I with ab325797 at 1/2000 (0.241 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : no staining on mouse kidney (PMID : 25760323).

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