Immunofluorescent analysis of PAX6 (green) in human B3 lens epithelial cells (left panel) and negative control U-2 OS cells (right panel). The cells were fixed with formaldehyde for 15 minutes, permeabilized with 0.1% Triton X-100 in TBS for 10 minutes, and blocked with 1% Blocker BSA in PBS (Product # 37525) for 15 minutes, all at room temperature. Cells were stained with a PAX6 monoclonal antibody (Product # MA1-109) in 1% Blocker BSA in PBS (Product # 37525) at a dilution of 1:100 for 1 hour at room temperature, and then incubated with a DyLight 488-conjugated goat anti-mouse IgG secondary antibody (Product # 35502) at a dilution of 1:250 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 ToxInsight Instrument at 20X magnification.
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Purified internal fragment of human recombinant PAX6 expressed in E. coli.|
|Storage buffer||PBS with 1mg/ml BSA, 30% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:100|
|Immunohistochemistry (Paraffin) (IHC (P))||1:50-1:200|
|Western Blot (WB)||1:500-1:2000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Western blot analysis of MA1-109 detects an ~47 kDa protein in whole cell lysates from early passage human B3 lens epithelial cells and in mouse eye lysate. An additional unknown band at ~36 kDa is also detected in B3 cell lysates. This antibody will detect additional nonspecific bands in lysates with very low basal PAX6 expression. Specificity of the antibody was confirmed in B3 and mouse eye lysates (positive controls) compared to U-2 OS human osteosarcoma and NIH/3T3 mouse embryonic fibroblasts (negative controls).
MA1-109- can be used for immunofluoresence and flow cytometry analysis of Pax6 in human neural stem cells.
PAX6 is a member of the paired box (PAX) family of transcription factors. Members of this gene family typically contain a paired box domain, an octapeptide, and a paired-type homeodomain. PAX6 has important functions in the development of the eye, nose, central nervous system and pancreas. Mutations in PAX6 are known to cause ocular disorders such as aniridia and Peter's anomaly. Transcript variants encoding 3 isoforms have been found for this gene, including isoform 1, isoform 5a and isoform 3 (PAX6-5A,6).
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.