Abstract
A radioimmunoassay for the measurement of rat pancreatic polypeptide (RPP) in serum or plasma has been developed and characterized using a new guinea-pig anti-rat-PP antibody. The assay provides a high degree of sensitivity and lacks cross-reactivity (CR < 0.01%) to neuropeptide Y and peptide YY. It also does not interact with PPs of other species or peptide hormones namely, amylin, glucagon, human insulin, human-PP, human-proinsulin, rat C-peptide and rat insulin. The assay employs synthetic rat PP as standards from concentrations of 21-2100 pg/ml (i.e., 5-500 pM) and produces a sensitivity limit of 19 pg/ml (4.5 pM) PP at ± 3 S.D. The intra- and interassay % coefficient of variations are 6.4% and 5.9%, respectively. The % recovery of RPP added to rat serum samples ranges from 98% to 103%. Assay of serum volumes ranging from 25 μl to 100 μl does not significantly alter the expected RPP level. The migration patterns of rat serum PP and that of a synthetic RPP are identical by Sephadex G-50 chromatographic analysis. The mean values of fasting and a 2 h post-feeding plasma RPP levels in normal rats are 40 ± 2 and 80 ± 10 pg/ml (9.5 pM and 19.0 pM), respectively. Rat-PP release during insulin induced hypoglycemia in conscious rats rises from 38 ± 5 pg/ml to 261 ± 34 pg/ml (9.0 to 62.1 pM, P < 0.005) by 30 min. Additionally, the antibody used in this study cross-reacts well with mouse-PP as determined by linear serum dilution curves, thus making it useful in the measurement of murine-PP. In conclusion, we have developed and validated a sensitive and specific rat-PP assay. This assay provides a new tool for the reliable measurement of PP in physiologic studies using rat and mouse animal models.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 59-69 |
| Number of pages | 11 |
| Journal | Regulatory Peptides |
| Volume | 37 |
| Issue number | 1 |
| DOIs | |
| State | Published - Jan 2 1992 |
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Keywords
- Pancreatic polypeptide
- Rat
- Specific radioimmunoassay
ASJC Scopus subject areas
- Biochemistry
- Endocrinology
- Physiology
- Neuroscience(all)
Cite this
The characterization of radioimmunoassay for rat pancreatic polypeptide in serum. / Akpan, Jones O.; Havel, Peter J; Parry, Susan J.; Shalwitz, Robert A.; Gingerich, Ronald L.
In: Regulatory Peptides, Vol. 37, No. 1, 02.01.1992, p. 59-69.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - The characterization of radioimmunoassay for rat pancreatic polypeptide in serum
AU - Akpan, Jones O.
AU - Havel, Peter J
AU - Parry, Susan J.
AU - Shalwitz, Robert A.
AU - Gingerich, Ronald L.
PY - 1992/1/2
Y1 - 1992/1/2
N2 - A radioimmunoassay for the measurement of rat pancreatic polypeptide (RPP) in serum or plasma has been developed and characterized using a new guinea-pig anti-rat-PP antibody. The assay provides a high degree of sensitivity and lacks cross-reactivity (CR < 0.01%) to neuropeptide Y and peptide YY. It also does not interact with PPs of other species or peptide hormones namely, amylin, glucagon, human insulin, human-PP, human-proinsulin, rat C-peptide and rat insulin. The assay employs synthetic rat PP as standards from concentrations of 21-2100 pg/ml (i.e., 5-500 pM) and produces a sensitivity limit of 19 pg/ml (4.5 pM) PP at ± 3 S.D. The intra- and interassay % coefficient of variations are 6.4% and 5.9%, respectively. The % recovery of RPP added to rat serum samples ranges from 98% to 103%. Assay of serum volumes ranging from 25 μl to 100 μl does not significantly alter the expected RPP level. The migration patterns of rat serum PP and that of a synthetic RPP are identical by Sephadex G-50 chromatographic analysis. The mean values of fasting and a 2 h post-feeding plasma RPP levels in normal rats are 40 ± 2 and 80 ± 10 pg/ml (9.5 pM and 19.0 pM), respectively. Rat-PP release during insulin induced hypoglycemia in conscious rats rises from 38 ± 5 pg/ml to 261 ± 34 pg/ml (9.0 to 62.1 pM, P < 0.005) by 30 min. Additionally, the antibody used in this study cross-reacts well with mouse-PP as determined by linear serum dilution curves, thus making it useful in the measurement of murine-PP. In conclusion, we have developed and validated a sensitive and specific rat-PP assay. This assay provides a new tool for the reliable measurement of PP in physiologic studies using rat and mouse animal models.
AB - A radioimmunoassay for the measurement of rat pancreatic polypeptide (RPP) in serum or plasma has been developed and characterized using a new guinea-pig anti-rat-PP antibody. The assay provides a high degree of sensitivity and lacks cross-reactivity (CR < 0.01%) to neuropeptide Y and peptide YY. It also does not interact with PPs of other species or peptide hormones namely, amylin, glucagon, human insulin, human-PP, human-proinsulin, rat C-peptide and rat insulin. The assay employs synthetic rat PP as standards from concentrations of 21-2100 pg/ml (i.e., 5-500 pM) and produces a sensitivity limit of 19 pg/ml (4.5 pM) PP at ± 3 S.D. The intra- and interassay % coefficient of variations are 6.4% and 5.9%, respectively. The % recovery of RPP added to rat serum samples ranges from 98% to 103%. Assay of serum volumes ranging from 25 μl to 100 μl does not significantly alter the expected RPP level. The migration patterns of rat serum PP and that of a synthetic RPP are identical by Sephadex G-50 chromatographic analysis. The mean values of fasting and a 2 h post-feeding plasma RPP levels in normal rats are 40 ± 2 and 80 ± 10 pg/ml (9.5 pM and 19.0 pM), respectively. Rat-PP release during insulin induced hypoglycemia in conscious rats rises from 38 ± 5 pg/ml to 261 ± 34 pg/ml (9.0 to 62.1 pM, P < 0.005) by 30 min. Additionally, the antibody used in this study cross-reacts well with mouse-PP as determined by linear serum dilution curves, thus making it useful in the measurement of murine-PP. In conclusion, we have developed and validated a sensitive and specific rat-PP assay. This assay provides a new tool for the reliable measurement of PP in physiologic studies using rat and mouse animal models.
KW - Pancreatic polypeptide
KW - Rat
KW - Specific radioimmunoassay
UR - http://www.scopus.com/inward/record.url?scp=0026531103&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0026531103&partnerID=8YFLogxK
U2 - 10.1016/0167-0115(92)90064-2
DO - 10.1016/0167-0115(92)90064-2
M3 - Article
VL - 37
SP - 59
EP - 69
JO - Regulatory Peptides
JF - Regulatory Peptides
SN - 0167-0115
IS - 1
ER -