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Immunofluorescence analysis of PAX6 was done on 70% confluent log phase U-87MG cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with PAX6 Rabbit Polyclonal Antibody (426600) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing nuclear localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Host / Isotype||Rabbit / IgG|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:100|
|Immunocytochemistry (ICC)||2-3 µg/mL|
|Immunofluorescence (IF)||2-3 µg/mL|
|Immunohistochemistry (Paraffin) (IHC (P))||1:100|
|Western Blot (WB)||1-2 µg/mL|
* 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.
42-6600 has been successfully used in flow cytometry and immunofluorescence analysis of Pax6 in human neural stem cells derived from pluripotent stem cells and reacts with human samples.
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"e; 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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
AN2; Aniridia; Aniridia Type II Protein; Dickie's small eye; Keratitis; Keratitis); MGDA; oculorhombin; Paired box 6; paired box gene 6; Paired Box Gene 6 (Aniridia; paired box homeotic gene 6; paired box homeotic gene-6; paired box protein Pax-6; Paired Box Protein-6; small eye; WAGR
1500038E17Rik; AEY11; AN; AN2; D11S812E; Dey; FVH1; Gsfaey11; MGDA; Pax-6; PAX6; Sey; WAGR