Anti-EGFR Picoband® Antibody

Western blot analysis of EGFR using anti-EGFR antibody (A00023-2).
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 50ug of sample under reducing conditions.
Lane 1: rat liver tissue lysates,
Lane 2: mouse liver tissue lysates,
Lane 3: mouse spleen tissue lysates,
Lane 4: mouse lung tissue lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-EGFR antigen affinity purified polyclonal antibody (Catalog # A00023-2) 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 EGFR at approximately 180KD. The expected band size for EGFR is at 180KD.

RHBDF2 related functions were mediated by EGFR signaling pathway. a , b Phosphorylation of EGFR and PD-L1 protein level in 786-O and 769-P cells were detected by western blot. Significance testing of gray statistics was analyzed by two-way ANOVA. c Growth rate of the transplanted tumor in the control group and RHBDF2 knockdown group (data were presented as the mean ± SEM and subjected to two-way ANOVA for significance test). d Representative images of xenografts in nude mice. e Immunofluorescent staining of phosphorylation of EGFR in the graft sections (The horizontal line at the bottom right represents 50 microns). f The mean fluorescence intensity of the phosphorylation staining of EGFR in the graft sections (t-test). g The migratory ability testing of 786-O cells and 769-P cells after Gefitinib treatment. h Statistics of the cell migratory ability after Gefitinib treatment (two-way ANOVA). i The detection of pEGFR and PD-L1 level in 786-O and 769-P cells after Gefitinib treatment. j Gray statistics of pEGFR and PD-L1 level in 786-O and 769-P cells (two-way ANOVA, * p < 0.05, ** p < 0.01, *** p < 0.005, **** p < 0.001)
Index in PubMed under a CC BY license. PMID: 34736454

IHC analysis of EGFR using anti-EGFR antibody (A00023-2).
EGFR was detected in paraffin-embedded section of mouse liver tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-EGFR Antibody (A00023-2) 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 EGFR using anti-EGFR antibody (A00023-2).
EGFR was detected in paraffin-embedded section of rat liver tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-EGFR Antibody (A00023-2) 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.