Anti-Apolipoprotein A I antibody [EPR28749-505]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Apolipoprotein A I antibody [EPR28749-505] (AB323863)

Immunohistochemical analysis of paraffin-embedded Rat non-perfused fixed kidney tissue labeling Apolipoprotein A I with ab323863 at 1/2000 (0.256 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on plasma in rat non-perfused fixed kidney. The primary antibody was incubated 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).

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-Apolipoprotein A I antibody [EPR28749-505] (AB323863)

Immunohistochemical analysis of paraffin-embedded (A) Rat non-perfused fixed liver and (B) Rat perfused fixed liver tissue labeling Apolipoprotein A I with ab323863 at 1/2000 (0.256 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on plasma in (A) rat non-perfused fixed liver, negative on (B) rat perfused fixed liver. The primary antibody was incubated 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).

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

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Apolipoprotein A I antibody [EPR28749-505] (AB323863)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized McA-RH7777 (rat liver epithelial cell) cells labelling Apolipoprotein A I with ab323863 at 1/2000 (0.256 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2ug/ml) dilution (Green).

Confocal image showing cytoplasmic staining in McA-RH7777 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Magenta).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2ug/ml) dilution.

• WB

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Western blot - Anti-Apolipoprotein A I antibody [EPR28749-505] (AB323863)

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

The expression profile/molecular weight observed is consistent with that described in the literature (PMID : 23969698, 23409054).

Low expression : liver, skeletal muscle (HPA database); brain (PMID : 1478650); lung (PMID : 23409054).

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

All lanes:

Western blot - Anti-Apolipoprotein A I antibody [EPR28749-505] (ab323863) at 1/1000 dilution

Lane 1:

Rat serum lysate at 20 µg

Lane 2:

Rat plasma lysate at 20 µg

Lane 3:

Rat liver tissue lysate at 20 µg

Lane 4:

Rat brain tissue lysate at 20 µg

Lane 5:

Rat lung tissue lysate at 20 µg

Lane 6:

Rat skeletal muscle 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 1/100000 dilution

Observed band size: 28 kDa,36 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-Apolipoprotein A I antibody [EPR28749-505] (AB323863)

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

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

All lanes:

Western blot - Anti-Apolipoprotein A I antibody [EPR28749-505] (ab323863) at 1/1000 dilution

Lane 1:

McA-RH7777 (rat liver epithelial cell) whole cell lysate at 20 µg

Lane 2:

McA-RH7777 culture supernatant 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: 30 kDa,36 kDa

false

Exposure time: 180s