Immunofluorescence was performed on mouse E10.5 embryo trunk at the forelimb level. Embryos were fixed with 4%PFA in 0.1M phosphate buffer, pH 7.2 at 4C for 30 minutes, processed for cryoprotection in 20% sucrose, and embedded in OCT compound for cryosectioning at 10 um thickness. Following sectioning, tissues were washed in PBS with 0.1% Triton-X 100 (PBT), blocked in 1% goat serum in PBT for 30 minutes at room temperature, and then probed with a Pax3 polyclonal antibody (Product # PA1-107) at a concentration of 1ug/ml in blocking buffer overnight at 4C. Tissues were washed extensively with PBT, and detection was performed using an Alexa Fluor 568-conjugated goat anti-rabbit IgG secondary antibody (red) at a dilution of 1:1000 in blocking buffer. Nuclei (blue) were stained with DAPI. Note: Staining was observed in the dorsal neural tube (symmetric two halves on the left side of the image), the dermomyotome/myotome (middle streak between symmetric halves), and limb muscle progenitors (scattered cells to the right side of the image). Data supplied courtesy of the Innovator's Program.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Amino acids 301-442 of human PAX3|
|Storage buffer||PBS with 1mg/ml BSA, 30% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Immunohistochemistry (Frozen) (IHC (F))||1:100-1:1000|
|Western Blot (WB)||1:500-1:1000|
* 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.
|Immunohistochemistry (IHC)||See 1 publications below|
PA1-107 has successfully been used in Western blot for detecting overexpressed and recombinant human Pax3 protein, but may not be suitable for detecting endogenous Pax3.
By immunofluorescence, PA1-107 has been used to stain frozen embryonic mouse brain tissue.
PA1-107 has been validated for IP using overexpressed Pax3 protein.
Pax 3 (Paired Box 3) is a transcription factor that is involved in development of the peripheral nervous system, melanocytes, some vascular smooth muscle, and a number of other derivatives. It regulates neurogenesis in pre-migratory neural crest cells from the dorsal neural tube, and in myogenic progenitors in the presomitic mesoderm and the hypaxial somites. Alternative splicing results in isoforms having different C-termini.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
NOVA regulates Dcc alternative splicing during neuronal migration and axon guidance in the spinal cord.
PA1-107 was used in immunohistochemistry to analyze regulation of Dcc alternative splicing during neuronal migration and axon guidance in the spinal cord by NOVA
|Leggere JC,Saito Y,Darnell RB,Tessier-Lavigne M,Junge HJ,Chen Z||eLife (5:null)||2016|