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In vitro assay for ACTH-releasing activity using ACTH radioimmunoassay. ACTH releasing activities by various drugs

Journal Article:

Abstract

This report deals with an in vitro assay of ACTH releasing activity utilizing pituitary incubation combined with ACTH radioimmunoassay. Half of a rat pituitary was preincubated in 2ml Krebs Ringer bicarbonate buffer containing 0.2% glucose and 0.25% BSA (KRBG-BSA) for 1.5 hr (45 min x 2). The medium was replaced by 1 ml KRBG-BSA and incubated for 30 min. Then the medium was again replaced by 1 ml KRBG-BSA or KRBG-BSA containing test materials and incubated for another 30 min. The amount of ACTH assayed by radioimmunoassay in the 2nd 30 min incubation was compared with that in the 1st 30 min incubation, and the result was expressed as a percentage. In the ACTH radioimmunoassay, anti-ACTH serum was diluted to 1:1,500-3,000. The /sup 125/I-..cap alpha../sup 1 -24/ACTH-antibody system was not affected by lysine-vasopressin (LVP), arginine-vasopressin (AVP), rat's pituitary LH, GH or prolactin. Human /sup 1 -39/ACTH was used as the ACTH standard. The dilution curve of the incubation medium was parallel to the standard curve. Reproducibility of immunoassayable ACTH within-assay was 174 +- 5.0 pg/tube (CV=2.9%). A log dose-relationship was observed between the amounts of stalk median eminence extracts (SME; NIAMDD) added to the incubation medium and its ACTH releasing  More>>
Authors:
Hashimoto, K; Takahara, J; Hosogi, H; Ofuji, N; Yasuhara, T [1] 
  1. Okayama Univ. (Japan). School of Medicine
Publication Date:
Feb 01, 1976
Product Type:
Journal Article
Reference Number:
AIX-08-283762; EDB-77-038517
Resource Relation:
Journal Name: Nippon Naibumpi Gakkai Zasshi; (Japan); Journal Volume: 52:2
Subject:
62 RADIOLOGY AND NUCLEAR MEDICINE; ACTH; RADIOIMMUNOASSAY; CALIBRATION; DILUTION; IMMUNE SERUMS; INCUBATION; IODINE 125; RADIOPHARMACEUTICALS; BETA DECAY RADIOISOTOPES; DAYS LIVING RADIOISOTOPES; DRUGS; ELECTRON CAPTURE RADIOISOTOPES; HORMONES; INTERMEDIATE MASS NUCLEI; IODINE ISOTOPES; ISOTOPE APPLICATIONS; ISOTOPES; LABELLED COMPOUNDS; NUCLEI; ODD-EVEN NUCLEI; PEPTIDE HORMONES; PITUITARY HORMONES; RADIOACTIVE MATERIALS; RADIOASSAY; RADIOISOTOPES; TRACER TECHNIQUES; 550601* - Medicine- Unsealed Radionuclides in Diagnostics
OSTI ID:
7324112
Country of Origin:
Japan
Language:
Japanese
Other Identifying Numbers:
Journal ID: CODEN: NNGZA
Submitting Site:
INIS
Size:
Pages: 114-124
Announcement Date:

Journal Article:

Citation Formats

Hashimoto, K, Takahara, J, Hosogi, H, Ofuji, N, and Yasuhara, T. In vitro assay for ACTH-releasing activity using ACTH radioimmunoassay. ACTH releasing activities by various drugs. Japan: N. p., 1976. Web.
Hashimoto, K, Takahara, J, Hosogi, H, Ofuji, N, & Yasuhara, T. In vitro assay for ACTH-releasing activity using ACTH radioimmunoassay. ACTH releasing activities by various drugs. Japan.
Hashimoto, K, Takahara, J, Hosogi, H, Ofuji, N, and Yasuhara, T. 1976. "In vitro assay for ACTH-releasing activity using ACTH radioimmunoassay. ACTH releasing activities by various drugs." Japan.
@misc{etde_7324112,
title = {In vitro assay for ACTH-releasing activity using ACTH radioimmunoassay. ACTH releasing activities by various drugs}
author = {Hashimoto, K, Takahara, J, Hosogi, H, Ofuji, N, and Yasuhara, T}
abstractNote = {This report deals with an in vitro assay of ACTH releasing activity utilizing pituitary incubation combined with ACTH radioimmunoassay. Half of a rat pituitary was preincubated in 2ml Krebs Ringer bicarbonate buffer containing 0.2% glucose and 0.25% BSA (KRBG-BSA) for 1.5 hr (45 min x 2). The medium was replaced by 1 ml KRBG-BSA and incubated for 30 min. Then the medium was again replaced by 1 ml KRBG-BSA or KRBG-BSA containing test materials and incubated for another 30 min. The amount of ACTH assayed by radioimmunoassay in the 2nd 30 min incubation was compared with that in the 1st 30 min incubation, and the result was expressed as a percentage. In the ACTH radioimmunoassay, anti-ACTH serum was diluted to 1:1,500-3,000. The /sup 125/I-..cap alpha../sup 1 -24/ACTH-antibody system was not affected by lysine-vasopressin (LVP), arginine-vasopressin (AVP), rat's pituitary LH, GH or prolactin. Human /sup 1 -39/ACTH was used as the ACTH standard. The dilution curve of the incubation medium was parallel to the standard curve. Reproducibility of immunoassayable ACTH within-assay was 174 +- 5.0 pg/tube (CV=2.9%). A log dose-relationship was observed between the amounts of stalk median eminence extracts (SME; NIAMDD) added to the incubation medium and its ACTH releasing activities. The sensitivity of this assay method was at least 0.1 SME or 10 mU of LVP and AVP. Using this method, it was found that LVP, AVP, norepinephrine (100 ng/ml--200 ng/ml) and 5-hydroxytryptophane (1 ..mu..g/ml) had ACTH releasing activities, but LH-RH, TRH, glucagon, dopamine, phentolamine, propranolol, haloperidol, prostaglandin E/sub 1/ and indomethacin did not affect the release of ACTH.}
journal = {Nippon Naibumpi Gakkai Zasshi; (Japan)}
volume = {52:2}
journal type = {AC}
place = {Japan}
year = {1976}
month = {Feb}
}