Radioimmunoassay for α-human and rat atrial natriuretic polypeptide

Nakao, Kazuwa; Sugawara, Akira; Morii, Narito; Sakamoto, Makoto; Suda, Michio; Soneda, J; Ban, Toshihiko; Kihara, Masahiro; Yamori, Yukio; Shimokura, Masanori; Kiso, Yoshiaki; Imura, Hiroo

doi:10.1016/0006-291x(84)91030-1

Cited by 144 publications

(64 citation statements)

References 15 publications

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“…Plasma ANP and cyclic guanosine monophosphate (cGMP) levels were determined by radioimmunoassay. 22 Before clamping and at the end of clamping, fractional excretion of sodium (FE Na ) was calculated by (urinary sodium × serum creatinine)/(serum sodium × urinary creatinine).…”

Section: Measurementsmentioning

confidence: 99%

Correlation of sodium-related factors with insulin sensitivity in young, lean, male offspring of hypertensive and normotensive subjects

et al. 2001

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Pioneer studies have proposed that multiple metabolic abnormalities, such as insulin resistance, increased Na + -H + exchanger activity and abnormal intracellular calcium homeostasis, are frequently associated with a subset of essential hypertensive patients with low plasma renin activity (PRA). However, it is unclear whether insulin resistance is related to the low renin status in the very early phase of genetical hypertension. Besides, there is controversy on the subject of the in vivo effect of acute hyperinsulinaemia on sodiumrelated factors. We investigated the relationship between sodium-related parameters and insulin sensitivity, and the effects of euglycaemic hyperinsulinaemia on cyclic guanosine monophosphate (cGMP) and atrial natriuretic peptide (ANP) levels in 17 young, lean, normotensive male subjects, who displayed extreme predis-

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Section: Measurementsmentioning

confidence: 99%

Correlation of sodium-related factors with insulin sensitivity in young, lean, male offspring of hypertensive and normotensive subjects

et al. 2001

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“…1 The ANP level was measured by RIA that recognizes the C-terminal sequence of immunoreactive ANP-(99-126). 3 …”

Section: Radioimmunoassaymentioning

confidence: 99%

Rat brain natriuretic peptide. Isolation from rat heart and tissue distribution.

et al. 1990

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We have isolated a cardiac natriuretic peptide of 5,000 d from atrial tissues from 500 rats and determined its amino acid sequence. The 5,000 d atrial natriuretic factor was elucidated to be a 45 amino acid peptide with the sequence of S-Q-D-S-A-F-R-I-Q-E-R-L-R-N-S-K-M-A-H-S-S-S-C-F-G-Q-K-I-D-R-I-G-A-V-S-R-L-G-C-D-G-L-R-L-F by sequencing the native peptideand its lysyl endopeptidase digests. The sequence of this peptide was identical to the amino acid sequence (51-95) of the rat brain natriuretic peptide precursor deduced from the complementary DNA (cDNA) sequence. The cardiac natriuretic peptide with a molecular weight of 5,000, or rat brain natriuretic peptide, was identified as the major storage form and as the sole secretory form derived from the brain natriuretic peptide precursor in the rat heart The rat brain natriuretic peptide level in the atrium was 3.68±0.61 /ig/g, which represents about 4% of that of atrial natriuretic factor. Rat brain natriuretic peptide was also detected in the ventricle. The ratio of brain natriuretic peptide to atrial natriuretic peptide in the ventricle was approximately 30% and much higher than that in the atrium. Rat brain natriuretic peptide, however, was not detectable in the brain. We conclude that the 5,000 d cardiac natriuretic peptide is rat brain natriuretic peptide with 45 amino acids derived from the brain natriuretic peptide precursor and is secreted from the rat heart as a novel cardiac hormone. (Hypertension 1990;15:774-778) B y using radioimmunoassay (RIA) for the ring structure of isoatriaJ natriuretic peptide (iso-ANP), which is the highly conserved sequence of natriuretic peptides, and radioreceptor assay for atrial natriuretic peptides (ANP), we have already demonstrated the presence of a considerable amount of a novel cardiac natriuretic peptide with a molecular weight of 5,000 in the rat atrium, which is distinct from iso-ANP and rat atrial natriuretic factor-(99-126) [rat ANP-(99-126)] isolated previously.

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“…6 Adult female BALB/c mice (Shizuoka Animal Center, Hamamatsu, Japan) were initially immunized by combined subcutaneous and intraperitoneal injections with the conjugate containing 30 /Ag of a-hANP emulsified in complete Freund's adjuvant (Difco Laboratories, Detroit, MI, USA). They were boosted in the same manner 3 weeks later and then bled to test for the presence of antibody by RIA with 125 I-a-hANP.…”

Section: Materials and Methods Preparation Of Monoclonal Antibodymentioning

confidence: 99%

“…6 In brief, a-hANP was radioiodinated by the chloramine-T method. The specific activity of l25 I-a-hANP ranged from 400 to 800 pCi/fig.…”

Section: Radioimmunoassaymentioning

confidence: 99%

“…2 -5 Using a polyclonal rabbit antiserum against a-ANP-(17-28) (ANF-[115-126]), we have established a specific radioimmunoassay (RIA) for ANP that recognizes a-human ANP (a-hANP or human ANF- ) and a-rat ANP (a-rANP or rat ANF-[99-126]) equally. 6 With the aid of this RIA, we have demonstrated that ANP is released from the heart and circulates in the body as a hormone 7 -8 and that ANP exists not only in the atrium but in the extraatrial tissues, including the brain, 9 spinal cord, 10 kidney," and lung. 12 This antiserum against a-ANP was also used for immunohistochemical studies.…”

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confidence: 99%

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A monoclonal antibody to alpha-human atrial natriuretic polypeptide.

et al. 1988

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SUMMARY A monoclonal antibody to o-human atrial natriuretic polypeptide (a-hANP), KY-ANP-I, hasbeen produced by fusion of a nonproducmg mouse myeloma cell tine, X63-Ag8.653, with spleen ceDs from BALB/c mice immunized with synthetic a-hANP conjugated to bovine thyrogtobuhn using the carbodiimide coupling procedure. Hybrklomas were screened for antibody production by rarJiolmmunoassay using culture media and l2S I-a-hANP. They were cloned by the llmtHng dilution technique, expanded hi culture, and injected mtraperHoneaDy into BALB/c mice. The obtained antibody belonged to the immunogiobunn G, subclass. Analysis by a Scatchard plot revealed a high affinity for a-hANP, with an association constant of 3.1 x 10" M~'. With this monoclonal antibody, a specific radioimmunoassay for a-hANP has been established. The antibody in mouse ascites was available for radioimmunoassay at a final dilution of 1:10*. Values of IC| 0 and IC<, in this radioimmunoassay were 3 and 30 fmol/tube, respectively. The radioimmunoassay showed a cross-reactivity of 0.9% with a-rat ANP. a-hANP-<8-22) and a-ANP-(l-6) exhibited less cross-reactivity than a-rat ANP on a molar basis. There was no cross-reaction with a-ANP-(17-28). Thus, the recognized epitope must be located in the N-terminal half of the ring structure of a-hANP including Met 12 residue. This radioimmunoassay could detect y-hANP and 0-hANP as well as a-hANP. The monoclonal antibody was also useful for unmunohistochemical studies. ANP-positive cells were finely stained hi the human atrium using the avidin-btotin-peroxidase complex technique. These results indicate that this monoclonal antibody to a-hANP will become a powerful tool for investigating the physiological and pathophvsiok>gkal significance of ANP. uretic polypeptides (ANPs), also known as atrial natriuretic factor (ANF), was isolated from human and rat atrial tissues and implicated in the control of water and electrolyte balance and blood pressure. kidney," and lung. 12 This antiserum against a-ANP was also used for immunohistochemical studies.

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Radioimmunoassay for α-human and rat atrial natriuretic polypeptide

Cited by 144 publications

References 15 publications

Correlation of sodium-related factors with insulin sensitivity in young, lean, male offspring of hypertensive and normotensive subjects

Correlation of sodium-related factors with insulin sensitivity in young, lean, male offspring of hypertensive and normotensive subjects

Rat brain natriuretic peptide. Isolation from rat heart and tissue distribution.

A monoclonal antibody to alpha-human atrial natriuretic polypeptide.

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