ab23248 detects ~50-55kDa band in rat sciatic nerve, rat and mouse brain lysate. IHC detection with ab23248 in mouse DRG shows a primarily nuclear localisation of the protein rather than the expected cytoplasmic staining (See Bao and Wang references).
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration. PubMed: 22366958
Use a concentration of 0.34 µg/ml.
Use a concentration of 1 µg/ml. Detects a band of approximately 52 kDa.
NRG1 type III is a sensory and motor neuron-derived factor isoform of Neuregulin 1, specifically expressed in the nervous system: spinal cord motor neurons, dorsal root ganglion neurons, and brain. NRG1 type III is the predominant isoform expressed in sensory and motor neurons.
The Nrg1 gene encodes more than 15 transmembrane and secreted protein isoforms, generated by alternative promoter usage and mRNA splicing. Nrg1 subtypes I through III share the epidermal growth factor-like signaling domain and are defined by different amino-termini. The N-terminal cysteine-rich domain (CRD), as found in the sensory and motor neuronderived factor (SMDF), defines Nrg1 type III, which in embryonic development is responsible for survival of Schwann cell precursors.
Immunohistochemistry (Frozen sections) - Anti-NRG1 type III antibody (ab23248)This image is courtesy of David Bennett, Kings College London, United Kingdom
ab23248 detecting NRG1 Type III in mouse L4/5 dorsal root ganglion (DRG) tissue (perfused with 4% paraformaldehyde and cryoprotected overnight, cut on a cryostat at 15µm). Tissue was incubated in blocking serum (normal goat serum) for 1hr, washed 3XPBS before primary antibody incubation in PBS with triton (0.1%) overnight (ab23248 was used at 1/800 = 0.34µg/ml). Secondary antibody used was goat anti-rabbit Alexa 546 (1/1000). The image shown is taken with a X20 objective and shows mostly nuclear NRG1 Type III staining with some cytomplasmic staining (nuclear counterstain with DAPI confirmed this - not shown).
Western blot - Anti-NRG1 type III antibody (ab23248)
All lanes : Anti-NRG1 type III antibody (ab23248) at 1 µg/ml
Lane 1 : Mouse Sciatic Nerve Lysate Lane 2 : Mouse Brain Lane 3 : Brain (Rat) Tissue Lysate (ab7942)
Lysates/proteins at 20 µg per lane.
Secondary All lanes : Rabbit IgG secondary antibody (ab28446) at 1/10000 dilution
ab23248 picks up an ~50kDa band in WB when tested on rat sciatic nerve, rat brain or mouse brain. This band size concurrs with an entry for NRG1 in both Expasy and NCBI databases (Q3URY7). Carroll et al., (J Neurosci. 1997 Mar 1;17(5):1642-59) also detect a 50kDa band in dorsal root ganglion and spinal cord. However, it is surprising that we do not detect more bands; it is likely that ab23248 does not pick up all NRG1 type III isoforms or that there is very restricted expression in the tissues tested here.
Yue Y et al. Low-intensity pulsed ultrasound upregulates pro-myelination indicators of Schwann cells enhanced by co-culture with adipose-derived stem cells. Cell Prolif49:720-728 (2016).
Read more (PubMed: 27625295) »
Sango K et al. Myelination in coculture of established neuronal and Schwann cell lines. Histochem Cell Biol137:829-39 (2012).
Read more (PubMed: 22366958) »