Neuroglycan C (NGC; also CSPG5 and CALEB) is a 120-160kD member of the neuregulin family of proteins. Depending upon its expression profile, it may be a glycoprotein oft 120kD, or a chondroitin sulfate (CS) proteoglycan of 140-160kD. NGC is expressed in nervous tissue and is found on retinal ganglion cells, cerebellar Purkinje cells and hippocampal neurons. Mature human NGC is a 536 amino acid (aa) type I transmembrane glycoprotein. It possesses a 393 aa extracellular region (aa 31-423) that contains a Ser117 CS attachment site and an EGF-like domain aa 371-413). There are at least two isoform variants. One shows a deletion of aa87-513, while a second shows an alternative start site at Met139 with the same deletion. Phosphorylation likely occurs on Ser249, and proteolysis generates a > 75kD soluble fragment. Over aa 31-420, human NGC shares 84% aa identity with mouse NGC.
Applications:
Suitable for Immunohistochemistry ELISA and Western Blot. Other applications not tested.
Recommended Dilution:
ELISA: 0.5-1ug/ml
Western Blot: 1ug/ml
Immunohistochemistry: 10ug/ml detects Neuroglycan C in sections of adult mouse cerebellum. Cells were fixed with PBS containing 4% paraformaldehyde for 20 minutes at RT and blocked
with PBS containing 10% normal donkey serum, 0.1% Triton X-100, and 1% BSA for 45 minutes at RT After blocking, cells were incubated with diluted primary antibody for 3 hours at RTfollowed by donkey anti-goat lgG at RT for 1 hour. Between each step, cells were washed with PBS containing 0.1% BSA.
Storage and Stability:
Lyophilized powder may be stored at 4 degrees C for short-term only. Reconstitute to nominal volume by adding sterile 40-50% glycerol and store at -20 degrees C. Reconstituted product is stable for 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
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