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Immunofluorescent analysis of PAX7 (green) in MCF7 and HeLa cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with a PAX7 polyclonal antibody (Product # PA1-117) at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with a DyLight 488-conjugated 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 ToxInsight Instrument at 20X magnification.
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Purified internal fragment of human recombinant PAX7 expressed in E. coli.|
|Storage buffer||PBS with 30% glycerol, 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Immunofluorescence (IF)||1:20 - 1:200|
|Immunoprecipitation (IP)||3 µg|
|Western Blot (WB)||1:1000 - 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 PA1-117 detects a prominent ~54 kDa protein in human HeLa, MCF7, rhabdomyosarcoma and skeletal muscle satellite cells and a weaker ~57 kDa protein in HeLa, rhabdomyosarcoma and skeletal muscle satellite cells. The ~57 kDa band seems to be enriched in immunoprecipitations from HeLa lysate, suggesting a bias for the larger isoform in ths application. Weak unknown bands of lower molecular weight and at ~110-130 kDa are also detected. Specificity of the antibody was also confirmed in Hela cells overexpressing full length PAX7.
PAX7 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. Pax7 is expressed in muscle satellite cells. The satellite cells are required for tissue repair and regeneration following muscle injury, and Pax7 expression is required to maintain survival and proliferation of postnatal satellite cells in adults. The chromosomal translocation t(1;13)(p36;q14) with FOXO1 in alveolar rhabdomyosarcomas, results in PAX7-FOXO1 fusion protein which acts as a transcriptional activator. Alternative splicing yields 2 isoforms, PAX7-A (57 kDa) and PAX7-B (54 kDa) (PMID: 15527970).
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
HUP1; paired box 7; paired box gene 7; paired box homeotic gene 7; paired box protein Pax-7; paired box transcription factor PAX7; paired domain gene 7; PAX7 transcriptional factor; PAX7A; PAX7B; RMS2
HUP1; Pax-7; PAX7; PAX7B; RMS2