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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: PC-3, Protein loading quantity: 20 µg, Exposure time: 60 s, Predicted MW: 68 kDa, Observed MW: 75 kDa. |
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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 orb1499364. |
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ICC staining of BMAL1 in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (orb1499364, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1000 dilution. The nuclear counter stain is DAPI (blue). |
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Tissue: Mouse hippocampus, Section type: Formalin fixed & Paraffin embedded section, Retrieval method: High temperature and high pressure, Retrieval buffer: Tris/EDTA buffer, pH9.0 Primary ab Dilution: 1:1000, Primary ab incubation condition: 1 hour at room temperature, Secondary ab: SP Kit (Rabbit), Counter stain: Hematoxylin (Blue), Comment: Color brown is the positive signal for orb1499364. |
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Tissue: Mouse pancreas, Section type: Formalin fixed & Paraffin embedded section, Retrieval method: High temperature and high pressure, Retrieval buffer: Tris/EDTA buffer, pH9.0 Primary ab Dilution: 1:1000, Primary ab incubation condition: 1 hour at room temperature, Secondary ab: SP Kit (Rabbit), Counter stain: Hematoxylin (Blue), Comment: Color brown is the positive signal for orb1499364. |
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Western blot analysis of BMAL1 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (orb1499364, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5000 dilution was used for 1 hour at room temperature. Positive control: Lane 1: Hela cell lysate, Lane 2: NIH/3T3 cell lysate, Lane 3: Rat brain tissue lysate, Lane 4: Mouse spleen tissue lysate. |