Anti-Apolipoprotein A I antibody [EP1368Y]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Apolipoprotein A I antibody [EP1368Y] (AB52945)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human liver tissue sections labeling Apolipoprotein A I with purified ab52945 at 1/100 dilution (1.95 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Apolipoprotein A I antibody [EP1368Y] (AB52945)

Intracellular Flow Cytometry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling Apolipoprotein A I with purified ab52945 at 1/20 dilution (10μg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluorr^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Apolipoprotein A I antibody [EP1368Y] (AB52945)

Immunocytochemistry/ Immunofluorescence analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling Apolipoprotein A I with purified ab52945 at 1/250 dilution (0.8 μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at 1/200 (2.5 μg/ml) dilution. Goat anti rabbit IgG (Alexa Fluor^® 488 ab150077) was used as the secondary antibody at 1/1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Apolipoprotein A I antibody [EP1368Y] (AB52945)

ab52945 (unpurified) at 1/100 dilution staining Apolipoprotein A I in human liver by Immunohistochemistry Paraffin embedded tissue.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Apolipoprotein A I antibody [EP1368Y] (AB52945)

ab52945 (unpurified) at 1/100 dilution staining Apolipoprotein A I in HEPG2 cells by Immunofluorescence.

• IP

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Immunoprecipitation - Anti-Apolipoprotein A I antibody [EP1368Y] (AB52945)

ab52945 (purified) at 1/20 dilution (1ug) immunoprecipitating Apolipoprotein A I in Human fetal liver lysates.
Lane 1 : Human fetal liver lysates 10ug
Lane 2 (+) : ab52945 & Human fetal liver lysates
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab52945 in Human fetal liver lysates
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.
Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-Apolipoprotein A I antibody [EP1368Y] (ab52945)

Predicted band size: 30 kDa

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• WB

Lab

Western blot - Anti-Apolipoprotein A I antibody [EP1368Y] (AB52945)

This blot was produced using 4-20% SDS-PAGE containing 15 μg of human liver lysate per lane at 150V for 1hr before being transferred onto a 0.45 μm PVDF membrane at 75V for 1hr. The membrane was then blocked for 1hr using 5% NFDM/TBST, then incubated with ab52945 (1/1000) at room temperature for 1hr. After being washed three times in TBST, the membrane was incubated with Peroxidase conjugated goat anti-rabbit IgG with minimal cross-reactivity with human IgG at 1/2000 dilution for 1hr at room temperature. The membrane was washed three times again. Then the signal was developed using the ECL technique. ab52945 was stored at a range of temperatures (+4°C, +22°C, +37°C) for 1 week before being tested in WB. The image shows the band intensity remains relatively constant across all storage temperatures, demonstrating that antibody activity is not affected.

All lanes:

Western blot - Anti-Apolipoprotein A I antibody [EP1368Y] (ab52945) at 1/1000 dilution

All lanes:

Human liver lysate at 15 µg with NDFM/TBST

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 1/2000 dilution

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Exposure time: 7s

• WB

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Western blot - Anti-Apolipoprotein A I antibody [EP1368Y] (AB52945)

All lanes:

Western blot - Anti-Apolipoprotein A I antibody [EP1368Y] (ab52945) at 1/20000 dilution

All lanes:

fetal liver lysate at 10 µg

Secondary

All lanes:

goat anti-rabbit HRP at 1/2000 dilution

Predicted band size: 30 kDa

Observed band size: 31 kDa

false

• I-ELISA

Lab

Indirect ELISA - Anti-Apolipoprotein A I antibody [EP1368Y] (AB52945)

ELISA analysis of Apolipoprotein A I recombinant protein at 1000 ng/mL with ab52945 at 1000~0ng/mL. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution was used as the secondary antibody.

• WB

Lab

Western blot - Anti-Apolipoprotein A I antibody [EP1368Y] (AB52945)

All lanes:

Western blot - Anti-Apolipoprotein A I antibody [EP1368Y] (ab52945) at 1/1000 dilution

All lanes:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates at 15 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

Predicted band size: 30 kDa

Observed band size: 31 kDa

false

• WB

Lab

Western blot - Anti-Apolipoprotein A I antibody [EP1368Y] (AB52945)

All lanes:

Western blot - Anti-Apolipoprotein A I antibody [EP1368Y] (ab52945) at 1/1000 dilution

All lanes:

Human liver lysates at 15 µg

Secondary

All lanes:

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

Predicted band size: 30 kDa

Observed band size: 31 kDa

false