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Bradykinin and Related Peptides

Mapping in Human Hepatoma Tissue or Rat Spinal Cord Tissue (H-009-01)

Tissue Sample

Human hepatoma tissue or Rat spinal cord tissue

Fixative

10% formalin

Embedding

Paraffin

Negative Control

No primary antibody

Pretreatment

Target retrieval 25 min (Steam)

Blocking

3% H2O2, 2% Normal Goat Serum

Primary Antibody

Anti-Bradykinin Serum (Catalog No.: H-009-01)

Optimal Dilution

1:500, 1 hour at RT

Secondary Antibody

Goat Anti-Rabbit IgG, Biotinylated (1:400), 30 min

Amplification

Streptavidin-HRP (Vector), 1:400, 30 min

Detection System

HRP

Substrate

DAB (Sigma), 3 min

Counterstained

Hematoxylin, 30 sec

Role of BK in nitric oxide (NO) formation on BPAEC

Role of BK in nitric oxide (NO) formation on BPAEC. BPAEC were preincubated with lisinopril (0.5mM) follow by incubation with 100nM HK and PK in HEPES-Tyrode gelatin buffer containing 2mM Ca2+, 1mM Mg2+, 8µM Zn2+, and 0.5mM lisinopril at 37C for 30min in the absence or presence of 100µM antipain, 5µM B2 BK receptor antagonist (B2 INHIBITOR, HOE 140, D-Arg-Arg-Pro-Hyp-Gly-Thi-Ser-D-Tic-Oic-Arg, Catalog No. 009-18, from Phoenix Pharmaceuticals), 5µM B1/B2 BK receptor antagonist (B1/B2 INHIBITOR, D-Arg-Arg-Pro-Hyp-Gly-Igl-Ser-D-Igl-Oic-Arg, Catalog No. 009-22, from Phoenix Pharmaceuticals), 0.2 to 0.5mM NG-nitro-L-arginine methyl ester (L-NAME), or 10 to 20µM NG-monomethyl-L-arginine (L-NMMA). At the end of the incubation, the wells were washed and the amount of NO formation was measured as indicated in the METHODS. Data are means ± SE of five determinations.
Zhao et al. Am J Physiol Heart Circ Physiol 2001 Apr;280(4):H1821-9

%bradykinin%


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