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 Dilution: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.
仕様
Size：100ug
Host：sheep
Source Antibody：human
Grade：Affinity Purified
Purity：Purified by immunoaffinity chromatography.
Form：Supplied as a lyophilized powder from PBS, 5% trehalose. Reconstitute with 500ul sterile PBS.
Specificity：Recognizes both the pro and mature forms of human Renin. In direct ELISAs and Western Blots, this antibody shows ~20% crossreactivity with rmRenin-1.
Isotype：IgG
Calc Applications Abbrev：E IHC IP N WB
Calc Crossreactivity：Hu
Immunogen：Recombinant protein corresponding to Leu24-Arg406 from human Renin, expressed in NS0. Species sequence homology: mouse 70%, canine 72%.
