Anti-pan SCN antibody [EPR25134-4]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pan SCN antibody [EPR25134-4] (AB300112)

Immunohistochemical analysis of paraffin-embedded human brain labeling SCN2A with ab300112 at 1/4000 dilution followed by a ready to use Leica DS9800 (BOND^™ Polymer Refine Detection).

Positive staining on human brain is observed. The section was incubated with ab300112 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Counterstained with Hematoxylin.

Secondary antibody only control : PBS was used instead of primary antibody followed by ready to use secondary antibody LeicaDS9800 (BOND^™ Polymer Refine Detection).

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pan SCN antibody [EPR25134-4] (AB300112)

Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue labeling SCN2A with ab300112 at 1/4000 dilution followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection).

Positive staining on mouse cerebellum is observed.

The section was incubated with ab300112 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

Counterstained with Hematoxylin.

Secondary antibody only control : PBS was used instead of primary antibody followed by ready to use secondary antibody LeicaDS9800 (BOND™ Polymer Refine Detection).

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

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-pan SCN antibody [EPR25134-4] (AB300112)

Fresh mouse cerebellum was fixed with 4% PFA and permeabilised with 0.2 % Triton X100.

Positive staining on mouse cerebellum.

ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed was used as a secondary antibody at 1/1000 dilution.

DAPI was used as a nuclear counterstain.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pan SCN antibody [EPR25134-4] (AB300112)

Immunohistochemical analysis of paraffin-embedded rat brain tissue labeling SCN2A with ab300112 at 1/4000 dilution followed by a ready to use Leica DS9800 (BOND^™ Polymer Refine Detection).

Positive staining on rat brain is observed.

The section was incubated with ab300112 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : PBS was used instead of primary antibody followed by ready to use secondary antibody LeicaDS9800 (BOND^™ Polymer Refine Detection).

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

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-pan SCN antibody [EPR25134-4] (AB300112)

Negative control. No staining on rat heart.

Fresh rat heart was fixed with 4% PFA and permeabilised with 0.2 % Triton X100. ab300112 was used as a primary antibody at 1/500 dilution.

ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed was used as a secondary antibody at 1/1000 dilution.

DAPI was used as a nuclear counterstain.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-pan SCN antibody [EPR25134-4] (AB300112)

Negative control : No staining on mouse heart.

Fresh mouse heart was fixed with 4% PFA and permeabilised with 0.2 % Triton X100. ab300112 was used as a primary antibody at 1/500 dilution.

ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed was used as a secondary antibody at 1/1000 dilution.

DAPI was used as a nuclear counter stain.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pan SCN antibody [EPR25134-4] (AB300112)

Immunohistochemical analysis of paraffin-embedded mouse brain labeling SCN2A with ab300112 at 1/4000 dilution followed by a ready to use Leica DS9800 (BOND^™ Polymer Refine Detection).

Positive staining on mouse brain in observed.

The section was incubated with ab300112 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Counterstained with Hematoxylin.

Secondary antibody only control : PBS was used instead of primary antibody followed by ready to use secondary antibody LeicaDS9800 (BOND^™ Polymer Refine Detection).

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

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-pan SCN antibody [EPR25134-4] (AB300112)

Positive staining on rat cerebellum.

Fresh rat cerebellum was fixed with 4% PFA and permeabilised with 0.2 % Triton X100. ab300112 was used as a primary antibody at 1/500 dilution.

ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed was used as a secondary antibody at 1/1000 dilution.

DAPI was used as a nuclear counterstain.

• IP

Supplier Data

Immunoprecipitation - Anti-pan SCN antibody [EPR25134-4] (AB300112)

Pan SCN was immunoprecipitated from 0.35 mg mouse brain tissue lysate with ab300112 at 1/30 dilution (2ug in 0.35mg lysates).

Western blot was performed on the immunoprecipitate using ab300112 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

All lanes:

Immunoprecipitation - Anti-pan SCN antibody [EPR25134-4] (ab300112) at 30 µg

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse brain tissue lysate at 1/30 IP at 20 µg

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab300112 in mouse brain tissue lysate at 20 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

false

Exposure time: 10s

• WB

Lab

Western blot - Anti-pan SCN antibody [EPR25134-4] (AB300112)

Exposure time : Lane 1-3 : 15 seconds; Lane 4 : 3 minutes.

This antibody does not cross-react with human SCN8A, SCN10A, and SCN11A.

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

All lanes:

Western blot - Anti-pan SCN antibody [EPR25134-4] (ab300112) at 1/1000 dilution

Lane 1:

HEK-293T cells transfected with a human SCN2A expression vector containing a his-tag, whole cell lysate at 20 µg

Lane 2:

HEK-293T cells transfected with a human SCN10A expression vector containing a his-tag, whole cell lysate at 20 µg

Lane 3:

HEK-293T cells transfected with a human SCN11A expression vector containing a his-tag, whole cell lysate at 20 µg

Lane 4:

HEK-293T cells transfected with a human SCN8A expression vector containing a his-tag, whole cell 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: 270 kDa

false

• WB

Lab

Western blot - Anti-pan SCN antibody [EPR25134-4] (AB300112)

Negative control : heart (PMID : 22081212).

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

All lanes:

Western blot - Anti-pan SCN antibody [EPR25134-4] (ab300112) 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 heart tissue 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: 270 kDa

false

Exposure time: 5.5s

• WB

Lab

Western blot - Anti-pan SCN antibody [EPR25134-4] (AB300112)

This antibody reacts with human SCN1A, SCN3A, and SCN9A but does not react with human SCN4A and SCN5A.

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

All lanes:

Western blot - Anti-pan SCN antibody [EPR25134-4] (ab300112) at 1/1000 dilution

Lane 1:

HEK-293T cells transfected with a human SCN1A fragment expression vector containing a his-tag, whole cell lysate at 20 µg

Lane 2:

HEK-293T cells transfected with a human SCN3A fragment expression vector containing a his-tag, whole cell lysate at 20 µg

Lane 3:

HEK-293T cells transfected with a human SCN4A fragment expression vector containing a his-tag, whole cell lysate at 20 µg

Lane 4:

HEK-293T cells transfected with a human SCN5A fragment expression vector containing a his-tag, whole cell lysate at 20 µg

Lane 5:

HEK-293T cells transfected with a human SCN9A fragment expression vector containing a his-tag, whole cell 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: 20-25 kDa

false

Exposure time: 26s

• WB

Lab

Western blot - Anti-pan SCN antibody [EPR25134-4] (AB300112)

Negative control : heart (PMID : 22081212).

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

All lanes:

Western blot - Anti-pan SCN antibody [EPR25134-4] (ab300112) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse hippocampus tissue lysate at 20 µg

Lane 3:

Mouse heart tissue lysate at 20 µg

Lane 4:

Rat brain tissue lysate at 20 µg

Lane 5:

Rat hippocampus tissue lysate at 20 µg

Lane 6:

Rat heart tissue 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: 270 kDa

false

Exposure time: 26s