* Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own experiment using appropriate negative and positive controls.
Antigen affinity chromatography
0.05% sodium azide
-20° C, Avoid Freeze/Thaw Cycles
Product Specific Information
PA1-801 detects PAX6 from human, mouse, rat, non-human primate and ovine samples. This antibody does not detect bovine PAX6 by Western blot.
PA1-801 has been successfully used in Western blot, immunofluorescence, and immunoprecipitation procedures. By Western blot, this antibody detects a ~47 kDa protein representing PAX6.
The PA1-801 immunogen is a synthetic peptide corresponding to residues C R(267) E E K L R N Q R R Q A S N T P S H I(285) of mouse PAX6. This sequence is completely conserved in humans, mice, frogs, and chickens. This peptide (Product # PEP-207) is available for use in neutralization and control experiments.
PAX genes encode nuclear transcription factors which are regarded as major controllers of developmental processes in both vertebrates and invertebrates. Mutations in murine PAX genes underlie three natural mouse alleles and several corresponding human syndromes (aniridia, foveal hypoplasia and Peters’ anomaly). Murine PAX genes have been shown to be proto-oncogenes. Furthermore, human PAX genes have recently been demonstrated to play an influential part in some common human cancers such as brain tumors and lymphomas. All PAX genes encode a DNA-binding domain termed the paired domain and in addition some also encode a second binding domain--the paired type homeobox. PAX6 is involved in the early development of the optical vesicle and has been shown to interact with Six3, another important visual development protein.
PAX6 Antibody (PA1-801) in WB PA1-801_1_07232010.jpg
Western Blot with anti-PAX6 Polyclonal Antibody (PA1-801)
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).
PAX6 Antibody (PA1-801) in IF PAX6-PA1-801-IF-1.jpg
Immunofluorescence with anti-PAX6 Polyclonal Antibody (PA1-801)
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.
PAX6 Antibody (PA1-801) in IP PAX6_PA1-801_IP.jpg
Immunoprecipitation with anti-PAX6 Polyclonal Antibody (PA1-801)
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).
(This product is for In Vitro experimental use only. Not for resale without express authorization.)