Anti-TRPV1 antibody [EPR25057-67]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRPV1 antibody [EPR25057-67] (AB305299)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human tissue labeling TRPV1 with ab305299 at 1/100 (5.14 µg/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Positive staining on (A) HEK-293T transfected with a TRVP1 His tag construct, no staining on (B) HEK-293T transfected with empty plasmid. The section was incubated with aab305299 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-TRPV1 antibody [EPR25057-67] (AB305299)

Immunofluorescent analysis of 4% Paraformaldehyde fixed mouse DRG primary neurons permeabilized using 0.1% Triton X-100 labelling TRPV1 with ab305299 (green) followed by ab150081 AlexaFluor^®488 preadsorbed Goat anti-Rabbit secondary. ab78078 anti-beta III Tubulin (mouse mAb) was used as counterstain (magenta) with ab150120 AlexaFluor^®594 Goat anti-Mouse as secondary. Nuclear DNA was labelled with DAPI (shown in blue). Image showing cytoplasmic staining in mouse DRG primary neurons was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRPV1 antibody [EPR25057-67] (AB305299)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling TRPV1 with ab305299 at 1/100 (5.14 µg/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Negative control : no staining on mouse cerebrum (PMID : 21451044).The section was incubated with ab305299 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRPV1 antibody [EPR25057-67] (AB305299)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling TRPV1 with ab305299 at 1/100 (5.14 µg/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Negative control : no staining on rat cerebrum (PMID : 21451044).The section was incubated with ab305299 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. 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-TRPV1 antibody [EPR25057-67] (AB305299)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat spleen (fresh) tissue labeling TRPV1 with ab305299 at 1/500 (1.028 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (Green). Low expression : confocal image showing no staining on rat spleen (PMID : 25409824). The nuclear counterstain was DAPI (Blue). The section was incubated with ab305299 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 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRPV1 antibody [EPR25057-67] (AB305299)

Immunohistochemical analysis of paraffin-embedded Mouse dorsal root ganglia tissue labeling TRPV1 with ab305299 at 1/100 (5.14 µg/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Positive staining on mouse dorsal root ganglia (PMID : 21451044).The section was incubated with ab305299 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. 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-TRPV1 antibody [EPR25057-67] (AB305299)

Immunohistochemical analysis of paraffin-embedded Rat dorsal root ganglia tissue labeling TRPV1 with ab305299 at 1/100 (5.14 µg/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Positive staining on Rat dorsal root ganglia (PMID : 28627595).The section was incubated with ab305299 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. 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-TRPV1 antibody [EPR25057-67] (AB305299)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat dorsal ganglion (fresh) tissue labeling TRPV1 with ab305299 at 1/500 (1.028 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (Green). Confocal image showing positive staining on rat dorsal ganglion. The nuclear counterstain was DAPI (Blue). The section was incubated with ab305299 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). is observed. 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 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-TRPV1 antibody [EPR25057-67] (AB305299)

Immunofluorescent analysis of 4% Paraformaldehyde fixed rat DRG primary neurons permeabilized using 0.1% Triton X-100 labelling TRPV1 with ab305299 (green) followed by ab150081 AlexaFluor^®488 preadsorbed Goat anti-Rabbit secondary. ab78078 anti-beta III Tubulin (mouse mAb) was used as counterstain (magenta) with ab150120 AlexaFluor^®594 Goat anti-Mouse as secondary. Nuclear DNA was labelled with DAPI (shown in blue). Image showing cytoplasmic staining in rat DRG primary neurons was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• IP

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Immunoprecipitation - Anti-TRPV1 antibody [EPR25057-67] (AB305299)

TRPV1 was immunoprecipitated from 0.35 mg Rat dorsal ganglion tissue lysate 10 μg with ab305299 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab305299. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Rat dorsal ganglion tissue lysate 10 μg

Lane 2 : ab305299 at 1/30 IP in Rat dorsal ganglion tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab305299 in Rat dorsal ganglion tissue lysate

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

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Immunoprecipitation - Anti-TRPV1 antibody [EPR25057-67] (ab305299) at 1/1000 dilution

Lane 1:

Rat dorsal ganglion tissue at 10 µg

Lane 2:

ab305299 at 1/30 IP in rat dorsal ganglion tissue lysate at 10 µg

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

Observed band size: 95 kDa

true

Exposure time: 3min

• WB

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Western blot - Anti-TRPV1 antibody [EPR25057-67] (AB305299)

Blocking and diluting buffer and concentration : 5% NFDM/TBST The molecular weight is consistent to what has been described in literature (PMID : 25395667) Negative control : brain, cerebellum (PMID : 11243859) Samples are non-boiled as boiling may cause protein aggregation. Exposure time : 114 seconds

All lanes:

Western blot - Anti-TRPV1 antibody [EPR25057-67] (ab305299) at 1/1000 dilution

Lane 1:

Rat dorsal galion tissue lysate at 20 µg

Lane 2:

Rat brain tissue lysate at 40 µg

Lane 3:

Human spinal cord at 20 µg

Lane 4:

Human cerebellum 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: 95 kDa

false

Exposure time: 114s

• WB

Lab

Western blot - Anti-TRPV1 antibody [EPR25057-67] (AB305299)

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

For Lane1, to minimize protein degradation, cell was lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.
Samples are non-boiled as boiling may cause protein aggregation.

All lanes:

Western blot - Anti-TRPV1 antibody [EPR25057-67] (ab305299) at 1/1000 dilution

Lane 1:

Rat fresh dorsal ganglion tissue lysate at 40 µg

Lane 2:

Rat dorsal ganglion 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 1/20000 dilution

Observed band size: 75 kDa,95 kDa

false

Exposure time: 180s