R1460-10
Renin
Size
£889.00
- SKU:
- R1460-10
- Purity:
- Purified by immunoaffinity chromatography.
- Storage Conditions:
- -20°C
- Supplier:
- United States Biological
- Host:
- Sheep
- Immunogen:
- Recombinant protein corresponding to Leu24-Arg406 from human Renin, expressed in NS0. Species sequence homology: mouse 70%, canine 72%.
- Format:
- Supplied as a lyophilized powder from PBS, 5% trehalose. Reconstitute with 500ul sterile PBS.
- Extra Details:
- Renin is a secreted, 44kD glycosylated member of the peptidase A1 family. It is an aspartyl protease that cleaves angiotensinogen to form angiotensin I. Human Renin is synthesized as a 47kD, 383 amino acid (aa) proform (aa # 24-406). In the kidney, pro-Renin is proteolytically cleaved after Arg66 to generate a mature enzyme. Activity is noted for both the proform and mature enzyme when bound to a cell surface receptor. Further processing may occur between Asn168 and Ser169 to generate 22kD, 102 aa N-terminal, and 18kD, 235 aa C-terminal fragments that are bioactive when associated. Applications: Suitable for use in ELISA, Western Blot, Immunohistochemistry, Immunoprecipitation and Neutralization. Other applications not tested. Recommended Dilutions: Western Blot: 0.1-0.2ug/ml with the appropriate secondary reagents to detect human Renin. The detection limit for rhRenin is approximately 5ng/lane under non-reducing and reducing conditions. Immunohistochemistry: 3-10ug/ml. Immunoprecipitation: Used to immunoprecipitate rhRenin from conditioned media of transfected NS0 cells. Neutralization: Used to inhibit/neutralize the enzymatic activity of rhRenin. The antibody was preincubated with the activated enzyme at different molar ratios at RT for 30 minutes. The enzyme was then assayed with a fluorogenic peptide substrate. 50% of the proteolytic activity was inhibited by the antibody at approximately 40ug/ml (IC50) under conditions in which the enzyme was present at 2ug/ml and the substrate concentration was 5uM. Considering the molecular masses of the enzyme (44kD) and the antibody (150 kD), IC50 was achieved at approximately 6:1 molar ratio of the antibody to the enzyme. Direct ELISA: 0.5-1.0ug/ml with the appropriate secondary reagents to detect human Renin. The detection limit for rhRenin is approximately 3ng/well. Optimal dilutions to be determined by the researcher. Storage and Stability: Lyophilized powder may be stored at -20°C. Stable for 12 months at -20°C. Reconstitute by adding sterile PBS. Aliquot and 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.
- Shipping Conditions:
- Blue Ice