Anti-IFNAR2 antibody [EPR30040-606]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IFNAR2 antibody [EPR30040-606] (AB325344)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling IFNAR2 with ab325344 at 1/500 (1.036 µg/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Low expression : nearly no staining on mouse skeletal muscle.

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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-IFNAR2 antibody [EPR30040-606] (AB325344)

Immunohistochemical analysis of paraffin-embedded (A) : J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage) and (B) : C2C12 (mouse myoblast) cell pellets labeling IFNAR2 with ab325344 at 1/500 (1.036 µg/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on J774A.1 cell pellet (image A), no staining on C2C12 cell pellet (image B) .

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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-IFNAR2 antibody [EPR30040-606] (AB325344)

Immunohistochemical analysis of paraffin-embedded Mouse breast cancer tissue labeling IFNAR2 with ab325344 at 1/500 (1.036 µg/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on mouse breast cancer.

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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-IFNAR2 antibody [EPR30040-606] (AB325344)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling IFNAR2 with ab325344 at 1/500 (1.036 µg/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on mouse spleen.

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

Lab

Western blot - Anti-IFNAR2 antibody [EPR30040-606] (AB325344)

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 8181059; PMID : 7759950).

Samples are non-boiled as boiling may cause protein aggregation.

In lane 1 and lane 3, the lysates were stored at -80°C prior to Western Blotting. In lane 2 and lane 4, To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

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

All lanes:

Western blot - Anti-IFNAR2 antibody [EPR30040-606] (ab325344) at 1/1000 dilution

Lane 1:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 2:

Neuro-2a (mouse neuroblastoma neuroblast) fresh whole cell lysate at 20 µg

Lane 3:

A20 (mouse reticulum sarcoma B lymphocyte) whole cell lysate at 20 µg

Lane 4:

A20 (mouse reticulum sarcoma B lymphocyte) fresh whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 40-100 kDa,36 kDa

false

Exposure time: 37s

• WB

Lab

Western blot - Anti-IFNAR2 antibody [EPR30040-606] (AB325344)

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

Low expression : skeletal muscle, brain

Samples are non-boiled as boiling may cause protein aggregation.

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

All lanes:

Western blot - Anti-IFNAR2 antibody [EPR30040-606] (ab325344) at 1/1000 dilution

Lane 1:

Mouse spleen tissue lysate at 20 µg

Lane 2:

Mouse lung tissue lysate at 20 µg

Lane 3:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 4:

Mouse brain tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 40-100 kDa,36 kDa

false

Exposure time: 37s

• WB

Lab

Western blot - Anti-IFNAR2 antibody [EPR30040-606] (AB325344)

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

Samples are non-boiled as boiling may cause protein aggregation.

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

All lanes:

Western blot - Anti-IFNAR2 antibody [EPR30040-606] (ab325344) at 1/1000 dilution

Lane 1:

Neuro-2a (mouse neuroblastoma neuroblast) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

Neuro-2a transfected with siRNA specifically targeting IFNAR2 whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 40-100 kDa,36 kDa

false

Exposure time: 26s