Goat Anti-HAPLN4 (Hyaluronan and Proteoglycan Link Protein 4, Brain Link Protein 2)
HAPLN4 (hyaluronan and proteoglycan link protein 4; also Bral2) is a 40-42kD secreted member of the Link module superfamily. It is secreted by select neurons, particularly of the cerebellum, where it participates in ECM stability. Here it may link hyaluronic acid with resident proteoglycans. Mature HAPLN4 is 383 amino acids (aa) in length. It contains one lg-like C2-type domain (aa 46-161) and two link domains (aa 163-365). One potential alternate start site exists 36 aa upstream of the standard start site. Human and mouse HAPLN4 share 91% aa sequence identity.
Applications
Western Blot: This antibody can be used at 0.1-0.2ug/ml with the appropriate secondary reagents to detect human HAPLN4. The detection limit for rhHAPLN4 is approximately 10 ng/lane under non-reducing and reducing conditions. Immunoprecipitation: This antibody has been used to immunoprecipitate rhHAPLN4 from conditioned media of transfected NS0 cells. Immunocytochemistry: This antibody has been used at a concentration of 10ug/ml to detect HAPLN4 on SHSY5Y human neuroblastoma cells. Cells were fixed with PBS containing 4% paraformaldehyde for 20 minutes at room temperature and blocked with PBS containing 10% normal donkey serum, 0.1% Triton X-100, and 1% BSA for 45 minutes at room temperature. After blocking, cells were incubated with diluted primary antibody overnight at 4° C followed by NorthernLightsTM 557-coupled donkey anti-goat IgG at room temperature in the dark for one hour. Between each step, cells were washed with PBS containing 0.1% BSA. Optimal dilutions should be deterhined by each laboratory for each application.
Storage and Stability
Lyophilized and reconstituted products are stable for 12 months after receipt at -20°C. Reconstitute with sterile PBS. Aliquot to avoid repeated freezing and thawing. Store at -20°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.
Immunogen
NS0-derived rhHAPLN4
Form
Supplied as a lyophilized powder from PBS, 5% trehalose. Reconstitute with 500ul sterile PBS.
Purity
Purified by immunoaffinity chromatography.
Specificity
Selected for its ability to recognize human HAPLN4 in the applications listed. In Western Blots, this antibody shows less than 1% crossreactivity with rhHAPLN1.