Anti-ProDynorphin antibody [EPR28632-48]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ProDynorphin antibody [EPR28632-48] (AB318966)

Immunohistochemical analysis of paraffin-embedded Mouse striatum tissue labeling ProDynorphin with ab318966 at 1/2000 (0.252 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in mouse striatum.
The section was incubated with ab318966 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

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Immunohistochemistry (Frozen sections) - Anti-ProDynorphin antibody [EPR28632-48] (AB318966)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat dorsomedial hypothalamic nucleus (perfused fixed) tissue labeling ProDynorphin with ab318966 at 1/50 (10.2 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Confocal image showing positive staining on rat dorsomedial hypothalamic nucleus (DMH). The nuclear counterstain was DAPI (Blue). The section was incubated with ab318966 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) preadsorbedat 1/1000 2 ug/mL dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ProDynorphin antibody [EPR28632-48] (AB318966)

Immunohistochemical analysis of paraffin-embedded Rat striatum tissue labeling ProDynorphin with ab318966 at 1/2000 (0.252 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in rat striatum.
The section was incubated with ab318966 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-ProDynorphin antibody [EPR28632-48] (AB318966)

Immunohistochemical analysis of paraffin-embedded Mouse pituitary tissue labeling ProDynorphin with ab318966 at 1/2000 (0.252 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in mouse pituitary.
The section was incubated with ab318966 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-ProDynorphin antibody [EPR28632-48] (AB318966)

Immunohistochemical analysis of paraffin-embedded Rat pituitary tissue labeling ProDynorphin with ab318966 at 1/2000 (0.252 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in rat pituitary.
The section was incubated with ab318966 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-ProDynorphin antibody [EPR28632-48] (AB318966)

Immunohistochemical analysis of paraffin-embedded Mouse hippocampus tissue labeling ProDynorphin with ab318966 at 1/2000 (0.252 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in mouse hippocampus.
The section was incubated with ab318966 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-ProDynorphin antibody [EPR28632-48] (AB318966)

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling ProDynorphin with ab318966 at 1/2000 (0.252 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining in rat liver (PMID : 3864164).
The section was incubated with ab318966 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-ProDynorphin antibody [EPR28632-48] (AB318966)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling ProDynorphin with ab318966 at 1/2000 (0.252 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining in mouse liver.
The section was incubated with ab318966 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

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-ProDynorphin antibody [EPR28632-48] (AB318966)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat liver (fresh frozen) tissue labeling ProDynorphin with ab318966 at 1/50 (10.06 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Negative control : confocal ima ge showing no staining on rat liver. The nuclear counterstain was DAPI (Blue). The section was incubated with ab318966 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) preadsorbedat 1/1000 2 ug/mL dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-ProDynorphin antibody [EPR28632-48] (AB318966)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse liver (fresh frozen) tissue labeling ProDynorphin with ab318966 at 1/50 (10.06 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Negative control : confocal ima ge showing no staining on mouse liver. The nuclear counterstain was DAPI (Blue). The section was incubated with ab318966 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) preadsorbedat 1/1000 2 ug/mL dilution.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-ProDynorphin antibody [EPR28632-48] (AB318966)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse hippocampus tissue staining ProDynorphin with ab318966 at a 1 : 5000 (0.1 ug/ml) dilution, SYNPR with ab314136 at 1 : 8000 (0.06 ug/ml) dilution and Wolframin with ab259362 at a 1 : 5000 (0.091 ug/ml) dilution followed by secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-ProDynorphin (magenta; Opal^™570), anti-SYNPR (green; Opal^™520) and anti-Wolframin (yellow; Opal^™690) on mouse hippocampus.
Panel B : anti-ProDynorphin staining the mossy fibers of CA3 in mouse hippocampus.
Panel C : anti-SYNPR staining the mossy fibers of CA3 in mouse hippocampus.
Panel D : anti-Wolframin staining the pyramidal neurons of CA1 in mouse hippocampus.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab318966, ab314136 and ab259362 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal^™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-ProDynorphin antibody [EPR28632-48] (AB318966)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse arcuate nucleus (perfused fixed) tissue labeling ProDynorphin with ab318966 at 1/50 (10.2 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Confocal image showing positive staining on mouse arcuate nucleus (ARC) of hypothalamus. The nuclear counterstain was DAPI (Blue). The section was incubated with ab318966 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) preadsorbedat 1/1000 2 ug/mL dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-ProDynorphin antibody [EPR28632-48] (AB318966)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat hippocampus (fresh frozen) tissue labeling ProDynorphin with ab318966 at 1/50 (10.06 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Panel A : merged staining of anti-ProDynorphin (ab318966, green), anti-NeuN (ab190565, grey) and anti-GFAP (ab201732, magenta) on rat hippocampus.
Panel B : anti-ProDynorphin stained on rat hippocampus.
Panel C : anti-NeuN stained in neurons of rat hippocampus.
Panel D : anti-GFAP stained in astrocytes of rat hippocampus.
The section was incubated in two rounds of staining : in the order of ab318966 and ab190565, ab201732 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) preadsorbedat 1/1000 2 ug/mL dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-ProDynorphin antibody [EPR28632-48] (AB318966)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse hippocampus (fresh frozen) tissue labeling ProDynorphin with ab318966 at 1/50 (10.06 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Panel A : merged staining of anti-ProDynorphin (ab318966, green), anti-NeuN (ab190565, grey) and anti-GFAP (ab201732, magenta) on mouse hippocampus.
Panel B : anti-ProDynorphin stained on mouse hippocampus.
Panel C : anti-NeuN stained in neurons of mouse hippocampus.
Panel D : anti-GFAP stained in astrocytes of mouse hippocampus.
The section was incubated in two rounds of staining : in the order of ab318966 and ab190565, ab201732 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) preadsorbedat 1/1000 2 ug/mL dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-ProDynorphin antibody [EPR28632-48] (AB318966)

ProDynorphin was immunoprecipitated from 0.35 mg Mouse pituitary tissue lysate with ab318966 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab318966 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse pituitary tissue lysate
Lane 2 : ab318966 IP in Mouse pituitary tissue lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab318966 in mouse pituitary tissue lysate

All lanes:

Immunoprecipitation - Anti-ProDynorphin antibody [EPR28632-48] (ab318966) at 1/30 dilution

All lanes:

Mouse pituitary tissue lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-ProDynorphin antibody [EPR28632-48] (AB318966)

Low expression : cerebellum (PMID : 16966485).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 16966485, PMID : 9712897).

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

All lanes:

Western blot - Anti-ProDynorphin antibody [EPR28632-48] (ab318966) at 1/1000 dilution

Lane 1:

Rat striatum tissue lysate at 20 µg

Lane 2:

Rat brain tissue lysate at 20 µg

Lane 3:

Rat cerebellum 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 1/100000 dilution

Observed band size: 18 kDa,28 kDa,36 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-ProDynorphin antibody [EPR28632-48] (AB318966)

Low expression : cerebellum (PMID : 16966485).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 16966485, PMID : 9712897).

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

All lanes:

Western blot - Anti-ProDynorphin antibody [EPR28632-48] (ab318966) at 1/1000 dilution

Lane 1:

Mouse striatum tissue lysate at 20 µg

Lane 2:

Mouse brain tissue lysate at 20 µg

Lane 3:

Mouse cerebellum 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 1/100000 dilution

Observed band size: 18 kDa,28 kDa,36 kDa

false

Exposure time: 81s

• WB

Supplier Data

Western blot - Anti-ProDynorphin antibody [EPR28632-48] (AB318966)

Negative control : liver(PMID : 3864164), kidney.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 16966485, PMID : 9712897).

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

All lanes:

Western blot - Anti-ProDynorphin antibody [EPR28632-48] (ab318966) at 1/1000 dilution

Lane 1:

Mouse pituitary tissue lysate at 20 µg

Lane 2:

Mouse kidney tissue lysate at 30 µg

Lane 3:

Mouse liver tissue lysate at 30 µg

Lane 4:

Rat pituitary tissue lysate at 20 µg

Lane 5:

Rat liver tissue lysate at 30 µg

Lane 6:

Rat kidney tissue lysate at 40 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: 18 kDa,28 kDa,36 kDa

false

Exposure time: 10s

• Dot

Supplier Data

Dot Blot - Anti-ProDynorphin antibody [EPR28632-48] (AB318966)

Dot blot analysis of ProDynorphin using ab318966 at 1 : 1000 (0.503 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

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

All lanes:

Dot Blot - Anti-ProDynorphin antibody [EPR28632-48] (ab318966) at 1/1000 dilution

Lane 1:

Mouse Proenkephalin-B full-length protein

Lane 2:

Mouse Proenkephalin-B pro-peptide

Lane 3:

Mouse Proenkephalin-B (Dynorphin A) peptide

Lane 4:

Mouse Proenkephalin-B (Alpha-neoendorphin) peptide

Lane 5:

Mouse Proenkephalin-B (Big dynorphin) peptide

Lane 6:

Mouse Proenkephalin-B peptide

Secondary

All lanes:

Dot 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

false

Exposure time: 180s