Anti-Apolipoprotein E4 antibody [EPR24181-64]

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-Apolipoprotein E4 antibody [EPR24181-64] (AB279714)

IHC image of Apolipoprotein E4 staining in a section of frozen human Alzheimer's disease brain* performed on a Leica BOND™ system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab279714, 0.05 μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-Apolipoprotein E4 antibody [EPR24181-64] (AB279714)

IHC image of Apolipoprotein E4 staining in a section of frozen normal human kidney* performed on a Leica BOND™ system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab279714, 0.05 μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Apolipoprotein E4 antibody [EPR24181-64] (AB279714)

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling Apolipoprotein E4 with ab279714 at 1/1000 dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human breast cancer. The section was incubated with ab279714 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) was used.

Heat mediated antigen retrieval 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-Apolipoprotein E4 antibody [EPR24181-64] (AB279714)

Immunohistochemical analysis of paraffin-embedded sections Panel A and D : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with APOE4 expression vector containing a myc-His-tag®
Panel B : HEK-293T transfected with APOE2 expression vector containing a myc-His-tag®. Panel C : HEK-293T transfected with APOE3 expression vector containing a myc-His-tag® labeling Apolipoprotein E4 with ab279714 at 1/1000 dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on (A) HEK-293T transfected with an APOE4 expression vector is observed. Almost no staining on (B) HEK-293T transfected with an APOE2 expression vector and (C) HEK-293T transfected with APOE3 expression vector. (D) Secondary antibody only control for (A). The section was incubated with ab279714 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) was used.

Heat mediated antigen retrieval 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-Apolipoprotein E4 antibody [EPR24181-64] (AB279714)

Immunohistochemical analysis of paraffin-embedded Human placenta (plasma) tissue labeling Apolipoprotein E4 with ab279714 at 1/1000 dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on the plasma in human placenta. (PMID : 26828652) The section was incubated with ab279714 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) was used.

Heat mediated antigen retrieval 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-Apolipoprotein E4 antibody [EPR24181-64] (AB279714)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Apolipoprotein E4 with ab279714 at 1/1000 dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human kidney. (PMID : 29692990 ) The section was incubated with ab279714 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) was used.

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

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Apolipoprotein E4 antibody [EPR24181-64] (AB279714)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T cells labelling Apolipoprotein E4 with ab279714 at 1/1000 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing strong cytoplasmic staining in 293T cells transfected with APOE4-Myc plasmid is observed. Myc-Tag Mouse mAb (Alexa Fluor® 647) antibody was used to counterstain transfected proteins at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Apolipoprotein E4 antibody [EPR24181-64] (AB279714)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T (Human embryonic kidney epithelial cell) transfected with myc-tagged Apolipoprotein E4 construct cells labeling Apolipoprotein E4 with ab279714 at 1/500 dilution (Right panel) compared with Rabbit monoclonal IgG Isotype Control (ab172730) (Left panel). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077)at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Apolipoprotein E4 antibody [EPR24181-64] (AB279714)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T cells labelling Apolipoprotein E4 with ab279714 at 1/500 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution. Negative control : Confocal image showing no staining in 293T cells transfected with APOE2-Myc or APOE3-Myc plasmid.. Myc-Tag Mouse mAb (Alexa Fluor® 647) antibody was used to counterstain transfected proteins at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Apolipoprotein E4 antibody [EPR24181-64] (AB279714)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed90% methanolpermeabilized 293T (Human embryonic kidney epithelial cell) transfected with myc-tagged Apolipoprotein E2 construct (Left)/ 293T transfected with myc tagged Apolipoprotein E3 construct (Right) labeling Apolipoprotein E4 with ab279714 at 1/500 dilution. Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077)at 1/2000 dilution was used as the secondary antibody.

• IP

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Immunoprecipitation - Anti-Apolipoprotein E4 antibody [EPR24181-64] (AB279714)

Apolipoprotein E4 was immunoprecipitated from 0.35 mg HEK-293T (human embryonic kidney) transfected with Apolipoprotein E4 expression vector containing a myc-His-tag® whole cell lysate 10 μg with ab279714 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab279714 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HEK-293T (human embryonic kidney) transfected with Apolipoprotein E4 expression vector containing a myc-His-tag® whole cell lysate 10 μg

Lane 2 : ab279746 IP in HEK-293T (human embryonic kidney) transfected with Apolipoprotein E4 expression vector containing a myc-His-tag® whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab279714 in HEK-293T transfected with Apolipoprotein E4 expression vector containing a myc-His-tag® whole cell lysate

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

Exposure time : 41 seconds

All lanes:

Immunoprecipitation - Anti-Apolipoprotein E4 antibody [EPR24181-64] (ab279714)

Predicted band size: 36 kDa

Observed band size: 40 kDa

false

• WB

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Western blot - Anti-Apolipoprotein E4 antibody [EPR24181-64] (AB279714)

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

Exposure time : 3 minutes

All lanes:

Western blot - Anti-Apolipoprotein E4 antibody [EPR24181-64] (ab279714) at 1/1000 dilution

All lanes:

Human plasma at 5 µL

Secondary

All lanes:

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

Predicted band size: 36 kDa

Observed band size: 36 kDa

false

• WB

Supplier Data

Western blot - Anti-Apolipoprotein E4 antibody [EPR24181-64] (AB279714)

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

All lanes:

Western blot - Anti-Apolipoprotein E4 antibody [EPR24181-64] (ab279714) at 1/1000 dilution

Lane 1:

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

Lane 2:

HEK-293T transfected with Apolipoprotein E2 expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

Lane 3:

HEK-293T transfected with Apolipoprotein E3 expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

Lane 4:

HEK-293T transfected with Apolipoprotein E4 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 1/50000 dilution

Predicted band size: 36 kDa

Observed band size: 40 kDa

false

Exposure time: 3min