Anti-Apolipoprotein A I/APOA1 Antibody Picoband®

Western blot analysis of APOA1 using anti-APOA1 antibody (PB9916).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human placenta tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-APOA1 antigen affinity purified polyclonal antibody (Catalog # PB9916) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for APOA1 at approximately 26 kDa. The expected band size for APOA1 is at 31 kDa.

IHC analysis of APOA1 using anti-APOA1 antibody (PB9916).
APOA1 was detected in a paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-APOA1 Antibody (PB9916) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

Comparison of the re-probed ability of PVDF membrane and NC membrane. The pooled sera proteins (3.0, 1.5, 0.7, 0.3 and 0.1 μg) were separated by 8% SDS-PAGE, and transferred to PVDF membranes (up) and NC membrane (down), respectively. ( a ) Staining with AAL and then re-probed with PHA-E; ( b ) staining with ApoA1 and then re-probing with IgG; ( c ) Staining with PHA-E and then re-probing with A2M; ( d ) staining with A2M and then re-probing with PHA-E. Band intensities were statistically analyzed (n = 3 individual experiments) and compared using Image Lab software (Bio-Rad Laboratories) and GraphPad Prism version 6. Pink bar, PVDF membrane; Blue bar, NC membrane. Band intensities were analyzed *Significantly different p < 0.05, ** p < 0.01. N.S., not significant. All values are means ± S.E. (error bars).
Index in PubMed under a CC BY license. PMID: 34103620

IHC analysis of APOA1 using anti-APOA1 antibody (PB9916).
APOA1 was detected in a paraffin-embedded section of human renal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-APOA1 Antibody (PB9916) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

Flow Cytometry analysis of HepG2 cells using anti-APOA1 antibody (PB9916).
Overlay histogram showing HepG2 cells stained with PB9916 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-APOA1 Antibody (PB9916, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.