Anti-S100A9 Antibody Picoband®

Western blot analysis of S100A9 using anti-S100A9 antibody (PB9678).
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: mouse spleen tissue lysates,
Lane 2: mouse lung 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-S100A9 antigen affinity purified polyclonal antibody (Catalog # PB9678) 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 S100A9 at approximately 13-15 kDa. The expected band size for S100A9 is at 13 kDa.

IHC analysis of S100A9 using anti-S100A9 antibody (PB9678).
S100A9 was detected in a paraffin-embedded section of mouse spleen 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-S100A9 Antibody (PB9678) 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.

IHC analysis of S100A9 using anti-S100A9 antibody (PB9678).
S100A9 was detected in a paraffin-embedded section of rat spleen 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-S100A9 Antibody (PB9678) 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.

IF analysis of S100A9 using anti-S100A9 antibody (PB9678).
S100A9 was detected in immunocytochemical section of HEPA1-6 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-S100A9 Antibody (PB9678) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Flow Cytometry analysis of HEPA1-6 cells using anti-S100A9 antibody (PB9678).
Overlay histogram showing HEPA1-6 cells stained with PB9678 (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-S100A9 Antibody (PB9678, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Validation of differential expression mRNAs and lncRNAs in spinal cord by real time RT-qPCR and Western blot analysis. (A) The differential expression mRNA levels were validated by qRT-PCR. (B) Five upregulated lncRNAs and five downregulated lncRNAs were validated by qRT-PCR. The levels of mRNAs and lncRNAs were normalized to GAPDH and expressed as fold of change compared to sham group. The results represent the mean± SEM of three independent experiments. *p < 0.05; **p < 0.01 compared with the sham group. Western blot analysis shown protein expression levels of Akt, P-Akt (C) , Bcl-2, Caspase-3 (D) , P2×7R, S100A9, Bax (E) . Each bar represents the mean ± SEM for at least 6 animals. *p< 0.05, **p< 0.01 vs Sham group.
Index in PubMed under a CC BY license. PMID: 28881824