Anti-Cytochrome P450 2E1 antibody [EPR28698-543]

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-Cytochrome P450 2E1 antibody [EPR28698-543] (AB324160)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human liver tissue staining Cytochrome P450 2E1 with ab324160 at a 1 : 2000 (0.262 ug/ml) dilution, ab202554 anti-COX IV used at 1 : 500 (0.228 ug/ml) dilution.

Panel A : merged staining of anti-CYP2E1 (green; Opal™520) and anti-COX IV (magenta; Opal™690) on human liver.
Panel B : anti-CYP2E1 staining hepatocytes in human liver.
Panel C : ant-COX IV staining hepatocytes in human liver.
Panel D : Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324160 and ab202554 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.

Nuclear counter stain with DAPI.

Heat mediated antigen retrieval 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-Cytochrome P450 2E1 antibody [EPR28698-543] (AB324160)

Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labeling Cytochrome P450 2E1 with ab324160 at 1/2000 (0.262 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : no staining on human cardiac muscle (PMID : 15576447). The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Slides were incubated 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 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-Cytochrome P450 2E1 antibody [EPR28698-543] (AB324160)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling Cytochrome P450 2E1 with ab324160 at 1/2000 (0.262 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : no staining on human spleen (PMID : 15576447). The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Slides were incubated 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 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-Cytochrome P450 2E1 antibody [EPR28698-543] (AB324160)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Cytochrome P450 2E1 with ab324160 at 1/2000 (0.262 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human liver (PMID : 25228302). The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Slides were incubated 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 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-Cytochrome P450 2E1 antibody [EPR28698-543] (AB324160)

Immunohistochemical analysis of paraffin-embedded Human hepatocellular carcinoma and adjacent liver tissue labeling Cytochrome P450 2E1 with ab324160 at 1/2000 (0.262 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining observed in adjacent human liver tissue, with reduced expression of CYP2E1 in hepatocellular carcinoma tissue. The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Slides were incubated 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 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-Cytochrome P450 2E1 antibody [EPR28698-543] (AB324160)

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling Cytochrome P450 2E1 with ab324160 at 1/2000 (0.262 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat liver (PMID : 35682741). The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Slides were incubated 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 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-Cytochrome P450 2E1 antibody [EPR28698-543] (AB324160)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Cytochrome P450 2E1 with ab324160 at 1/2000 (0.262 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse liver (PMID : 23914059). The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Slides were incubated 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-Cytochrome P450 2E1 antibody [EPR28698-543] (AB324160)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded rat liver tissue staining Cytochrome P450 2E1 with ab324160 at a 1 : 2000 (0.262 ug/ml) dilution, ab202554 anti-COX IV used at 1 : 500 (0.228 ug/ml) dilution.

Panel A : merged staining of anti-CYP2E1 (green; Opal™520) and anti-COX IV (magenta; Opal™690) on rat liver.
Panel B : anti-CYP2E1 staining hepatocytes in rat liver.
Panel C : ant-COX IV staining hepatocytes in rat liver.
Panel D : Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324160 and ab202554 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.

Nuclear counter stain with DAPI.

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

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-Cytochrome P450 2E1 antibody [EPR28698-543] (AB324160)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse liver tissue staining Cytochrome P450 2E1 with ab324160 at a 1 : 2000 (0.262 ug/ml) dilution, ab202554 anti-COX IV used at 1 : 500 (0.228 ug/ml) dilution.

Panel A : merged staining of anti-CYP2E1 (green; Opal™520) and anti-COX IV (magenta; Opal™690) on mouse liver.
Panel B : anti-CYP2E1 staining hepatocytes in mouse liver.
Panel C : ant-COX IV staining hepatocytes in mouse liver.
Panel D : Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324160 and ab202554 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.

Nuclear counter stain with DAPI.

Heat mediated antigen retrieval 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-Cytochrome P450 2E1 antibody [EPR28698-543] (AB324160)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling Cytochrome P450 2E1 with ab324160 at 1/2000 (0.262 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : no staining on rat spleen. The primary antibody was incubated for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Slides were incubated 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 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-Cytochrome P450 2E1 antibody [EPR28698-543] (AB324160)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Cytochrome P450 2E1 with ab324160 at 1/2000 (0.262 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : no 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. Slides were incubated 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• WB

Supplier Data

Western blot - Anti-Cytochrome P450 2E1 antibody [EPR28698-543] (AB324160)

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

Low expression : colon, spleen (PMID : 15576447).

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

All lanes:

Western blot - Anti-Cytochrome P450 2E1 antibody [EPR28698-543] (ab324160) at 1/1000 dilution

Lane 1:

Human liver tissue lysate at 20 µg

Lane 2:

Human colon tissue lysate at 20 µg

Lane 3:

Human spleen 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: 57 kDa,36 kDa

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Anti-Cytochrome P450 2E1 antibody [EPR28698-543] (AB324160)

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

Low expression : heart, spleen, colon (PMID : 15576447).

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

All lanes:

Western blot - Anti-Cytochrome P450 2E1 antibody [EPR28698-543] (ab324160) at 1/1000 dilution

Lane 1:

Mouse liver tissue lysate at 20 µg

Lane 2:

Mouse heart tissue lysate at 20 µg

Lane 3:

Mouse spleen tissue lysate at 20 µg

Lane 4:

Mouse colon tissue lysate at 20 µg

Lane 5:

Rat liver tissue lysate at 20 µg

Lane 6:

Rat heart tissue lysate at 20 µg

Lane 7:

Rat spleen tissue lysate at 20 µg

Lane 8:

Rat colon 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 1/20000 dilution

Observed band size: 57 kDa,36 kDa

false

Exposure time: 3s

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Cytochrome P450 2E1 antibody [EPR28698-543] (AB324160)

Immunofluorescent analysis of 4% Paraformaldehyde fixed 0.1% Triton X-100 permeabilized Mouse primary hepatocytes labelling Cytochrome P450 2E1 with ab324160 at 1/500 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 μg/ml) dilution (Green). Confocal image showing cytoplasmic staining in mouse primary hepatocyte (shown in green). The counterstain (ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594)) at 1/1000 (2 μg/ml) was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).

On image : -ve conrol 1, ab324160 at 1/500 (1.046 μg/ml) with ab150120 at 1/1000 (2μg/ml). -ve conrol 2, Anti- Human Serum Albumin at 1/500 (2μg/ml) with ab150081 at 1/1000 (2μg/ml).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Cytochrome P450 2E1 antibody [EPR28698-543] (AB324160)

Immunofluorescent analysis of 4% Paraformaldehyde fixed 0.1% Triton X-100 permeabilized Mouse Mouse splenocytes labelling Cytochrome P450 2E1 with ab324160 at 1/500 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 μg/ml) dilution (Green). Negative control : Confocal image showing no staining in mouse splenocytes. The counterstain (ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594)) at 1/1000 (2 μg/ml) was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).

On image : -ve conrol 1, ab324160 at 1/500 (1.046 μg/ml) with ab150120 at 1/1000 (2μg/ml). -ve conrol 2, Anti- Human Serum Albumin at 1/500 (2μg/ml) with ab150081 at 1/1000 (2μg/ml).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• WB

Supplier Data

Western blot - Anti-Cytochrome P450 2E1 antibody [EPR28698-543] (AB324160)

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

This antibody does not cross-react with overexpressed human Cytochrome P450 2C19 or Cytochrome P450 2C18 by western blot.

In Western blot, Anti-6X His tag® antibody [AD1.1.10] - (ab15149) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-Cytochrome P450 2E1 antibody [EPR28698-543] (ab324160) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag®, whole cell lysate at 20 µg

Lane 2:

293T transfected with a human Cytochrome P450 2E1 expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 3:

293T transfected with a human Cytochrome P450 2C19 expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 4:

293T transfected with a human Cytochrome P450 2C18 expression vector containing a myc-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 1/20000 dilution

Observed band size: 57 kDa,36 kDa,56 kDa

false

Exposure time: 10s