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- domestic rabbit polyclonal
reactivity: human, mouse, rat, domestic sheep
application: western blot, immunocytochemistry, immunoprecipitation
citations: 3
Western blot analysis of PAX6 was performed by loading 50ug of various human, mouse and non-human primate whole cell lysates per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with a PAX6 polyclonal antibody (Product # PA1-801) at a dilution of 1:1000 overnight at 4°C on a rocking platform, washed in TBS-0.1%Tween 20, and probed with a goat anti-rabbit IgG-HRP secondary antibody (Product # 31460) at a dilution of 1:20,000 for at least one hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34075).
Immunoprecipitation of PAX6 was performed on 293T cells. Antigen: antibody complexes were formed by incubating 500µg whole cell lysate with 5µg of PAX6 polyclonal antibody (Product # PA1-801) overnight on a rocking platform at 4°C. The immune complexes were captured on 50µl Protein A/G Plus Agarose (Product # 20423) , washed extensively, and eluted with 5X Lane Marker Reducing Sample Buffer (Product # 39000). Samples were resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membrane, and blocked with 5% BSA/TBS-0.1%Tween for at least 1 hour. The membrane was probed with a PAX6 monoclonal antibody (Product # PA1-801) at a dilution of 1:1000 overnight rotating at 4°C, washed in TBST, and probed with Clean-Blot IP Detection Reagent (Product # 21230) at a dilution of 1:1000 for at least one hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34075).
Immunofluorescent analysis of PAX6 (green) in All-Trans-Retinoic Acid (ATRA) treated (10µM, 24 hours) NCCIT cells. 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 (Product # 37525) for 15 minutes at room temperature. Cells were probed with a PAX6 polyclonal antibody (Product # PA1-801), at a dilution of 1:50 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat-anti-rabbit IgG secondary antibody (Product # 35552) at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DyLight 554 Phalloidin (Product # 21834), and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan at 20X magnification.
quantity: 100 ug
price: US 465.00
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gene information - domestic sheep PAX6
- description:paired box 6
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- PAX6 antibody
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