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For every force there is an opposing force that balances the overall effect. Such is the basic tenant of Yin and Yang. Adrenomedullin (ADM) eloquently fits this ancient wisdom. Hypotensive effects are elicited by fragments of ADM retaining the C-terminal with ring structure. Hypertensive effects are seen with N-terminal ADM fragments containing the ring structure, or with the ring structure alone. The hypotensive effects can be antagonized with C-terminal fragments lacking the ring structure. Nature has endowed ADM with the ability to regulate itself, however it remains to be seen if the Yin and Yang of ADM is utilized in vivo.
 
Catalog No.: WBK-010-10 
| Rat heart tissue |
| 10% formalin |
| paraffin |
| No primary antibody |
| Target retrieval 25 min (Steam) |
| 3% H2O2, 2% Normal Goat Serum |
| Anti-ADM (rat) Antibody (Catalog No.: H-010-08) |
| 1:200, 1 hour at RT |
| Goat Anti-Rabbit IgG, Biotinylated (1:400), 30 min |
| Streptavidin-HRP (Vector), 1:400, 30 min |
| HRP |
| DAB (Sigma), 3 min |
| Hematoxylin, 30 sec |
Accumulation of ADM in the culture medium of CAECs.  indicates spontaneous secretion of ADM from CAECs;  , ADM secretion from CAECs stimulated with sarafotoxin S6c (10-7 mol/L). *P<0.05 between groups. Hypertension. 1998;32:917-922
After incubation for the indicated time, the culture medium was removed and immediately acidified with acetic acid (final concentration, 1 mol/L), heated at 100°C for 10 minutes to inactivate intrinsic protease, and stored at -20°C until assay. Culture medium (1 mL) was extracted on C-18 Bond Elute cartridges and eluted with 75% methanol containing 1% trifluoroacetic acid. Concentrated eluates were then assayed using a specific and sensitive radioimmunoassay for ADM(1-52) (Phoenix Pharmaceuticals Inc) as previously described. Minimal detectable concentration for the assay is 1 pg per tube, and the half-maximal inhibition dose of radioiodinated ligand binding by ADM is 20 pg per tube. Recovery is 72%, and intra-assay and interassay variations are 10% and 12%, respectively.
Jougasaki M., et al. Hypertension. 1998;32:917-922
Immunocytochemistry-- Rabbit anti-rat ADM antibodies were from Phoenix Pharmaceuticals Inc. and were used at a 1/200 dilution. After fixation with 4% paraformaldehyde, cells were permeabilized with 0.2% Triton X-100 (v/v). Nonspecific binding sites were saturated using 4% normal goat serum (v/v). Anti-ADM antibodies were detected with a goat anti-rabbit antibody coupled to rhodamine (TRITC) (Jackson Immunoresearch).
Annie Ladoux and Christian Frelin. J. Biol. Chem., 2000, 275 (51), 39914-39919
ADM Prohormone Schematics
PAMP Sequence Comparison
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