Western blot analysis of beta 2 Adrenergic Receptor on zebrafish tissue lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (bsm-52880R, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:5,000 dilution was used for 1 hour at room temperature.

Blocking buffer: 5% NFDM/TBST Primary Ab dilution: 1:1000 Primary Ab incubation condition: lane 1: PTM-6016+Competitive peptides, lane 2: PTM- 6016, 2 hours at room temperature Secondary Ab: Goat Anti-Rabbit IgG H&L (HRP) Lysate: Mouse kidney Protein loading quantity: 20 μg Exposure time: 60 s Predicted MW: 46 kDa Observed MW: 46 kDa

Blocking buffer: 5% NFDM/TBST Primary Ab dilution: 1:2000 Primary Ab incubation condition: 2 hours at room temperature Secondary Ab: Goat Anti-Rabbit IgG H&L (HRP) Lysate: 1: A431, 2: MCF-7, 3: MDA-MB-231, 4: Rat kidney, 5: Mouse kidney Protein loading quantity: 20 μg Exposure time: 60 s Predicted MW: 46 kDa Observed MW: 46 kDa

Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-beta 2 Adrenergic Receptor antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (bsm-52880R, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemical analysis of paraffin-embedded mouse liver tissue using anti-beta 2 Adrenergic Receptor antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (bsm-52880R, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Cell line: A431 Fixation: 100% Ice-cold methanol Permeabilization: 0.1% TritonX-100 Primary Ab dilution: 1:100 Primary Ab incubation condition: 4°C overnight Secondary Ab: Goat Anti-Rabbit IgG Nuclear counter stain: DAPI (Blue) Comment: Color green is the positive signal for bsm-52880R

Cell line: MCF7 Fixation: 100% Ice-cold methanol Permeabilization: 0.1% TritonX-100 Primary Ab dilution: 1:100 Primary Ab incubation condition: 4°C overnight Secondary Ab: Goat Anti-Rabbit IgG Nuclear counter stain: DAPI (Blue) Comment: Color green is the positive signal for bsm-52880R

Cell line: SH-SY5Y Fixation: 100% Ice-cold methanol Permeabilization: 0.1% TritonX-100 Primary Ab dilution: 1:50 Primary Ab incubation condition: 4℃ overnight Secondary Ab: Goat Anti-Rabbit IgG Nuclear counter stain: DAPI (Blue) Comment: Color green is the positive signal for PTM-6016

Cell line: SH-SY5Y Fixation: 4% Paraformaldehyde Permeabilization: 90% Methanol Primary Ab dilution: 1:100 Secondary Ab: Goat Anti-Rabbit IgG Unlabelled control: The cell without incubation with primary antibody and secondary antibody (Black line). Isotype control: Rabbit monoclonal IgG (Blue line). Comment: Line red is the positive signal for bsm-52880R