Primary antibody notesab27303 is NRG1 Type I specific and therefore is predicted to detect the NRG1 beta Type I isoforms 6-8 (NRG1 Beta1, Beta1A, Beta2, Beta3) in addition to the NRG1 alpha isoforms 1-4.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml.
1/100 - 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Use a concentration of 1 µg/ml. Detects a band of approximately 51 kDa (predicted molecular weight: 70 kDa).
Use at an assay dependent concentration. PubMed: 21203579
FunctionDirect ligand for ERBB3 and ERBB4 tyrosine kinase receptors. Concomitantly recruits ERBB1 and ERBB2 coreceptors, resulting in ligand-stimulated tyrosine phosphorylation and activation of the ERBB receptors. The multiple isoforms perform diverse functions such as inducing growth and differentiation of epithelial, glial, neuronal, and skeletal muscle cells; inducing expression of acetylcholine receptor in synaptic vesicles during the formation of the neuromuscular junction; stimulating lobuloalveolar budding and milk production in the mammary gland and inducing differentiation of mammary tumor cells; stimulating Schwann cell proliferation; implication in the development of the myocardium such as trabeculation of the developing heart. Isoform 10 may play a role in motor and sensory neuron development.
Tissue specificityType I isoforms are the predominant forms expressed in the endocardium. Isoform alpha is expressed in breast, ovary, testis, prostate, heart, skeletal muscle, lung, placenta liver, kidney, salivary gland, small intestine and brain, but not in uterus, stomach, pancreas, and spleen. Isoform 3 is the predominant form in mesenchymal cells and in non-neuronal organs, whereas isoform 6 is the major neuronal form. Isoform 8 is expressed in spinal cord and brain. Isoform 9 is the major form in skeletal muscle cells; in the nervous system it is expressed in spinal cord and brain. Also detected in adult heart, placenta, lung, liver, kidney, and pancreas. Isoform 10 is expressed in nervous system: spinal cord motor neurons, dorsal root ganglion neurons, and brain. Predominant isoform expressed in sensory and motor neurons. Not detected in adult heart, placenta, lung, liver, skeletal muscle, kidney, and pancreas. Not expressed in fetal lung, liver and kidney. Type IV isoforms are brain-specific.
Involvement in diseaseNote=A chromosomal aberration involving NRG1 produces gamma-heregulin. Translocation t(8;11) with ODZ4. The translocation fuses the 5'-end of ODZ4 to NRG1 (isoform 8). The product of this translocation was first thought to be an alternatively spliced isoform. Gamma-heregulin is a soluble activating ligand for the ERBB2-ERBB3 receptor complex and acts as an autocrine growth factor in a specific breast cancer cell line (MDA-MB-175). Not detected in breast carcinoma samples, including ductal, lobular, medullary, and mucinous histological types, neither in other breast cancer cell lines.
Sequence similaritiesBelongs to the neuregulin family. Contains 1 EGF-like domain. Contains 1 Ig-like C2-type (immunoglobulin-like) domain.
Developmental stageDetectable at early embryonic ages. Isoform 10 is highly expressed in developing spinal motor neurons and in developing cranial nerve nuclei. Expression is maintained only in both adult motor neurons and dorsal root ganglion neurons. Type IV isoforms are expressed in fetal brain.
DomainThe cytoplasmic domain may be involved in the regulation of trafficking and proteolytic processing. Regulation of the proteolytic processing involves initial intracellular domain dimerization. ERBB receptor binding is elicited entirely by the EGF-like domain.
Post-translational modificationsProteolytic cleavage close to the plasma membrane on the external face leads to the release of the soluble growth factor form. N- and O-glycosylated. Extensive glycosylation precedes the proteolytic cleavage.
Cellular localizationSecreted; Cell membrane. Does not seem to be active; Membrane. May possess an internal uncleaved signal sequence; Nucleus. May be nuclear and Secreted. Has a signal peptide.
The predicted MW for Isoform 1 of this protein is ~70 kDa. The 51 kDa band observed in our data correlates with the predicted size of Isoform 3 (50.7 kDa). We believe the doublet is caused by the presence of a smaller processed form of Isoform 3.
The epitope for ab27303 is present in all the described isoforms of the protein.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NRG1 type I antibody (ab27303)This image is courtesy of Carl Hobbs, King's College London, United Kingdom
Immunohistochemistical (Formaldehyde/PFA-fixed paraffin-embedded sections) staining for NRG1 type I antibody (ab27303) on Zebrafish Tissue sections (Transverse section through whole body). This image shows specific staining for NRG1 Type 1 in the spinal cord. Antigen retrieval step: Heat mediated. Blocking step: 1% BSA for 10 mins at RT. Primary Antibody ab27303 incubated at 1/100 for 16 hours at RT. Secondary Antibody: Biotin labelled goat anti Rabbit IgG (1/300).
Immunocytochemistry/ Immunofluorescence - NRG1 type I antibody (ab27303)
ICC/IF image of ab27303 stained human HEK 293 cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab27303, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - NRG1 type I antibody (ab27303)
IHC image of NRG1 type 1 staining in human liver carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab27303, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
References for Anti-NRG1 type I antibody (ab27303)
This product has been referenced in:
Marques-Smith A et al. A Transient Translaminar GABAergic Interneuron Circuit Connects Thalamocortical Recipient Layers in Neonatal Somatosensory Cortex. Neuron89:536-49 (2016).
Read more (PubMed: 26844833) »
Pan P & Dobrowsky RT Differential expression of neuregulin-1 isoforms and downregulation of erbin are associated with Erb B2 receptor activation in diabetic peripheral neuropathy. Acta Neuropathol Commun1:39 (2013).
Read more (PubMed: 24252174) »