Anti-SYNPR antibody [EPR28139-74]

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-SYNPR antibody [EPR28139-74] (AB314136)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse liver (fresh) tissue labeling SYNPR with ab314136 at 1100 (5.08 ugml) 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 image showing no staining on mouse liver (PMID : 12974474). The nuclear counterstain was DAPI (Blue). The section was incubated with ab314136 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). The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-SYNPR antibody [EPR28139-74] (AB314136)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat liver (fresh) tissue labeling SYNPR with ab314136 at 1100 (5.08 ugml) 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 image showing no staining on rat liver (PMID : 12974474). The nuclear counterstain was DAPI (Blue). The section was incubated with ab314136 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). The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SYNPR antibody [EPR28139-74] (AB314136)

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling SYNPR with ab314136 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on mouse kidney (PMID : 2206533). The section was incubated with ab314136 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-SYNPR antibody [EPR28139-74] (AB314136)

Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling SYNPR with ab314136 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on rat liver. The section was incubated with ab314136 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-SYNPR antibody [EPR28139-74] (AB314136)

Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling SYNPR with ab314136 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on mouse liver (PMID : 2206533). The section was incubated with ab314136 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-SYNPR antibody [EPR28139-74] (AB314136)

Immunohistochemical analysis of paraffin-embedded rat hippocampus tissue labeling SYNPR with ab314136 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat hippocampus (PMID : 12271488). The section was incubated with ab314136 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-SYNPR antibody [EPR28139-74] (AB314136)

Immunohistochemical analysis of paraffin-embedded mouse hippocampus tissue labeling SYNPR with ab314136 at 1/2000 (0.254 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse hippocampus (PMID : 12271488). The section was incubated with ab314136 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-SYNPR antibody [EPR28139-74] (AB314136)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse hippocampus (fresh) tissue labeling SYNPR with ab314136 at 1100 (5.08 ugml) 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-SYNPR (ab314136, green), anti-Map2 (ab225315, red) on mouse hippocampus. Panel B : anti-SYNPR stained on mouse hippocampus. Panel C : anti-Map2 stained on mouse hippocampus. The section was incubated in two rounds of staining in the order of ab314136 and ab225315 for 1 hour 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). The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-SYNPR antibody [EPR28139-74] (AB314136)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat hippocampus (fresh) tissue labeling SYNPR with ab314136 at 1100 (5.08 ugml) 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-SYNPR (ab314136, green), anti-Map2 (ab225315, red) on rat hippocampus. Panel B : anti-SYNPR stained on rat hippocampus. Panel C : anti-Map2 stained on rat hippocampus. The section was incubated in two rounds of staining : in the order of ab314136 and ab225315 for 1 hour 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). The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-SYNPR antibody [EPR28139-74] (AB314136)

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

• mIHC

Lab

Multiplex immunohistochemistry - Anti-SYNPR antibody [EPR28139-74] (AB314136)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Rat hippocampus tissue staining SYNPR with ab314136 at a 1 : 8000 (0.06 ug/ml) dilution, BACE1 with ab263901 at 1 : 4000 (0.193 ug/ml) dilution and 5HT6 Receptor with ab315380 at a 1 : 1000 (0.504 ug/ml) dilution followed by secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-SYNPR (green; Opal™520), anti-BACE1 (magenta;Opal™570) and anti-5HT6 Receptor (yellow;Opal™690) on rat hippocampus.
Panel B : anti-SYNPR staining the mossy fibers in rat hippocampus.
Panel C : anti-BACE1 staining the mossy fibers in rat hippocampus.
Panel D : anti-5HT6 Receptor staining the pyramidal neurons in rat hippocampus.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab314136, ab263901 and ab315380 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.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-SYNPR antibody [EPR28139-74] (AB314136)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse hippocampus tissue staining SYNPR with ab314136 at a 1 : 8000 (0.06 ug/ml) dilution, BACE1 with ab263901 at 1 : 4000 (0.193 ug/ml) dilution and 5HT6 Receptor with ab315380 at a 1 : 1000 (0.504 ug/ml) dilution followed by secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-SYNPR (green; Opal™520), anti-BACE1 (magenta;Opal™570) and anti-5HT6 Receptor (yellow;Opal™690) on mouse hippocampus.
Panel B : anti-SYNPR staining the mossy fibers in mouse hippocampus.
Panel C : anti-BACE1 staining the mossy fibers in mouse hippocampus.
Panel D : anti-5HT6 Receptor staining the pyramidal neurons 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 ab314136, ab263901 and ab315380 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.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-SYNPR antibody [EPR28139-74] (AB314136)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse hippocampus tissue staining SYNPR with ab314136 at a 1 : 8000 (0.06 ug/ml) dilution, WFS1 with ab259362 at 1 : 5000 (0.091 ug/ml) dilution and 5HT6 Receptor with ab315380 at a 1 : 1000 (0.504 ug/ml) dilution followed by secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-SYNPR (yellow; Opal™520), anti-WFS1 (green;Opal™570) and anti-5HT6 Receptor (magenta;Opal™690) on mouse hippocampus.
Panel B : anti-SYNPR staining the mossy fibers in mouse hippocampus.
Panel C : anti-WFS1 staining the pyramidal neurons of CA1 in mouse hippocampus.
Panel D : anti-5HT6 Receptor staining the pyramidal neurons 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 ab314136, ab259362 and ab315380 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.

• IP

Supplier Data

Immunoprecipitation - Anti-SYNPR antibody [EPR28139-74] (AB314136)

SYNPR was immunoprecipitated from 0.35 mg Rat brain tissue lysate with ab314136 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314136 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : Rat brain tissue lysate Lane 2 : Rat brain tissue lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab314136 in rat brain tissue lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 8 seconds

All lanes:

Immunoprecipitation - Anti-SYNPR antibody [EPR28139-74] (ab314136) at 1/30 dilution

All lanes:

Rat brain 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: 8s

• IP

Supplier Data

Immunoprecipitation - Anti-SYNPR antibody [EPR28139-74] (AB314136)

SYNPR was immunoprecipitated from 0.35 mg Mouse brain tissue lysate with ab314136 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314136 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : Mouse brain tissue lysate Lane 2 : Mouse brain tissue lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab314136 in mouse brain tissue lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 8 seconds

All lanes:

Immunoprecipitation - Anti-SYNPR antibody [EPR28139-74] (ab314136) at 1/30 dilution

All lanes:

Mouse brain 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: 8s

• WB

Supplier Data

Western blot - Anti-SYNPR antibody [EPR28139-74] (AB314136)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : kidney (PMID : 12974474, 2206533), skeletal muscle (PMID : 12974474). The expression profile observed is consistent with what has been described in literature (PMID : 12974474, 2206533). In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution. Exposure time : 180 seconds

All lanes:

Western blot - Anti-SYNPR antibody [EPR28139-74] (ab314136) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

Human hypothalamus tissue lysate at 20 µg

Lane 3:

Human kidney tissue lysate at 20 µg

Lane 4:

Human skeletal muscle lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-SYNPR antibody [EPR28139-74] (AB314136)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : kidney, liver (PMID : 12974474, 2206533). The expression profile observed is consistent with what has been described in literature (PMID : 12974474, 2206533). In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution. Exposure time : Lane 1-5 : 114 seconds ; Lane 6-7 : 180 seconds.

All lanes:

Western blot - Anti-SYNPR antibody [EPR28139-74] (ab314136) at 1/1000 dilution

Lane 1:

Mouse hypothalamus tissue lysate at 20 µg

Lane 2:

Mouse cerebellum tissue lysate at 20 µg

Lane 3:

Mouse kidney tissue lysate at 20 µg

Lane 4:

Rat brain lysate at 20 µg

Lane 5:

Rat kidney lysate at 20 µg

Lane 6:

Rat hypothalamus lysate at 20 µg

Lane 7:

Rat liver 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: 36 kDa

false

• WB

Supplier Data

Western blot - Anti-SYNPR antibody [EPR28139-74] (AB314136)

Blocking and diluting buffer and concentration : 5% NFDM/TBST SYNPR detected a shift from 36kDa to 29kDa band after deglycosylation treatment. In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution. Exposure time : 92 seconds

All lanes:

Western blot - Anti-SYNPR antibody [EPR28139-74] (ab314136) at 1/1000 dilution

Lane 1:

Untreated mouse hippocampus tissue lysate at 20 µg

Lane 2:

Mouse hippocampus tissue lysate treated with Protein treated with Peptide:N-glycosidase F (PNGase F) at 20 µg

Lane 3:

Mouse brain 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: 36 kDa

false

Exposure time: 92s