Anti-Amphiregulin/Areg Antibody Picoband®

FCGR2B were up-regulated in hippocampus of DM mice. A qRT-PCR was performed to detect the expression of ALB, AREG and FCGR2B mRNA expression in hippocampus of mice. B Western blot was conducted to detect the ALB, AREG and FCGR2B protein expression in hippocampus of mice. C IHC assay was employed to examine the ALB, AREG and FCGR2B protein expression in hippocampus of mice. D IF staining was utilized to detect the expression of FCGR2B and NeuN in hippocampus of mice. E Western blot was performed to detect the SHC1 protein expression in hippocampus of mice. F IF staining was performed to detect the expression of SHC1 and NeuN in hippocampus of mice. G Western blot was used to detect the p-PI3K and p-AKT protein expression in hippocampus of mice. *** P < 0.001 Full size image
Index in PubMed under a CC BY license. PMID: 40537751

Western blot analysis of Amphiregulin using anti-Amphiregulin antibody (A01787-1).
Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: rat brain tissue lysates,
Lane 2: rat thymus tissue lysates,
Lane 3: rat spleen tissue lysates,
Lane 4: rat PC-12 whole cell lysates,
Lane 5: mouse brain tissue lysates,
Lane 6: mouse thymus tissue lysates,
Lane 7: mouse spleen tissue lysates,
Lane 8: mouse RAW264.7 whole cell 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-Amphiregulin antigen affinity purified polyclonal antibody (A01787-1) 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 (Catalog # BA1054) at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for Amphiregulin at approximately 45 kDa. The expected band size for Amphiregulin is at 28 kDa.

IHC analysis of Amphiregulin using anti-Amphiregulin antibody (A01787-1).
Amphiregulin was detected in a paraffin-embedded section of mouse brain 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 2 μg/ml rabbit anti-Amphiregulin Antibody (A01787-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of Amphiregulin using anti-Amphiregulin antibody (A01787-1).
Amphiregulin was detected in a paraffin-embedded section of rat brain 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 2 μg/ml rabbit anti-Amphiregulin Antibody (A01787-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

Flow Cytometry analysis of PC-12 cells using anti-Amphiregulin antibody (A01787-1).
Overlay histogram showing PC-12 cells stained with A01787-1 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-Amphiregulin Antibody (A01787-1, 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.