- Overview
- Data/Specifications
- Literature/Support
- How It Works
- Related Products
Overview
PTH (Parathyroid hormone, Parathormone, Parathyrin) is biosynthesized in the parathyroid gland as a pre-proparathyroid hormone, a larger molecular precursor consisting of 115 amino acids. Following sequential intracellular cleavage of a 25-amino acid sequence, preproparathyroid hormone is converted to an intermediate, a 90-amino acid polypetide, proparathyroid hormone. With additional proteolytic modification, proparathyroid hormone is then converted to parathyroid hormone, an 84 amino acid polypeptide. In healthy individuals, regulation of parathyroid hormone secretion normally occurs via a negative feedback action of serum calcium on the parathyroid glands. Intact PTH is biologically active and clears very rapidly from the circulation with a half-life of less than four minutes. PTH undergoes proteolysis in the parathyroid glands, but mostly peripherally, particularly in the liver but also in the kidneys and bone, to give N-terminal fragments and longer lived C-terminal and midregion fragments. In subjects with renal insufficiency, C-terminal and midregion PTH assays typically give elevated PTH results, as reflected by impaired renal clearance.
Data/Specifications
Species: human
Sample Type:serum, plasma
Sample Size:25uL
Standard Curve Range: 7 - 700 pg/mL
Sensitivity: 0.9 pg/mL
Assay Length: 3.5 hrs
Literature/Support
Product Insert:
Intact PTH ELISA (product insert, PDF)
Articles/Troublshooting:
ELISA Troubleshooting Guide
ELISA Data Reduction Guide
References:
Bauer, J. M., Mikušová, L., Verlaan, S., Bautmans, I., Brandt, K., Donini, L. M., ... & Luiking, Y. (2020). Safety and tolerability of 6-month supplementation with a vitamin D, calcium and leucine-enriched whey protein medical nutrition drink in sarcopenic older adults. Aging Clinical and Experimental Research, 1-14.
Cashman, K. D., Kenny, S., Kerry, J. P., Leenhardt, F., & Arendt, E. K. (2019). ‘Low-salt’bread as an important component of a pragmatic reduced-salt diet for lowering blood pressure in adults with elevated blood pressure.Nutrients,11(8), 1725.
Jennings, A., Cashman, K. D., Gillings, R., Cassidy, A., Tang, J., Fraser, W., ... & Pietruszka, B. (2018). A Mediterranean-like dietary pattern with vitamin D3 (10 µg/d) supplements reduced the rate of bone loss in older Europeans with osteoporosis at baseline: results of a 1-y randomized controlled trial.The American journal of clinical nutrition,108(3), 633-640.
O"Callaghan, K. M., Hennessy, Á., Hull, G. L., Healy, K., Ritz, C., Kenny, L. C., ... & Kiely, M. E. (2018). Estimation of the maternal vitamin D intake that maintains circulating 25-hydroxyvitamin D in late gestation at a concentration sufficient to keep umbilical cord sera≥ 25–30 nmol/L: a dose-response, double-blind, randomized placebo-controlled trial in pregnant women at northern latitude.The American journal of clinical nutrition.
Hemmingway, A., Kenny, L. C., Malvisi, L., & Kiely, M. E. (2018). Exploring the concept of functional vitamin D deficiency in pregnancy: impact of the interaction between 25-hydroxyvitamin D and parathyroid hormone on perinatal outcomes.The American journal of clinical nutrition,108(4), 821-829.
Carson, E. L., Pourshahidi, L. K., Madigan, S. M., Baldrick, F. R., Kelly, M. G., Laird, E., ... & Mulhern, M. S. (2018). Vitamin D status is associated with muscle strength and quality of life in patients with COPD: a seasonal prospective observation study.International journal of chronic obstructive pulmonary disease,13, 2613.
Hayes, A., Duffy, S., O’Grady, M., Jakobsen, J., Galvin, K., Teahan-Dillon, J., ... & Seamans, K. M. (2016). Vitamin D–enhanced eggs are protective of wintertime serum 25-hydroxyvitamin D in a randomized controlled trial of adults, 2.The American journal of clinical nutrition,104(3), 629-637.
O"Donovan, C. B., Walsh, M. C., Nugent, A. P., McNulty, B., Walton, J., Flynn, A., & Brennan, L. (2015). Use of metabotyping for the delivery of personalised nutrition. Molecular nutrition & food research, 59(3), 377-385.
Cashman, K. D., Hayes, A., O"Donovan, S. M., Zhang, J. Y., Kinsella, M., Galvin, K., ... & Seamans, K. M. (2014). Dietary calcium does not interact with vitamin D3 in terms of determining the response and catabolism of serum 25-hydroxyvitamin D during winter in older adults. The American journal of clinical nutrition, 99(6), 1414-1423.
Wang, J., Trentham-Dietz, A., Hemming, J. D., Hedman, C. J., & Sprague, B. L. (2013). Serum factors and clinical characteristics associated with serum E-Screen activity. Cancer Epidemiology Biomarkers & Prevention, 22(5), 962-971.
Muldowney, S., Lucey, A. J., Hill, T. R., Seamans, K. M., Taylor, N., Wallace, J. M., ... & Kiely, M. (2012). Incremental cholecalciferol supplementation up to 15 μg/d throughout winter at 51–55 N has no effect on biomarkers of cardiovascular risk in healthy young and older adults. The Journal of nutrition, 142(8), 1519-1525.
Birmingham, D. J., Hebert, L. A., Song, H., Noonan, W. T., Rovin, B. H., Nagaraja, H. N., & Yu, C. Y. (2012). Evidence that abnormally large seasonal declines in vitamin D status may trigger SLE flare in non-African Americans. Lupus, 21(8), 855-864.
Cashman, K. D., Seamans, K. M., Lucey, A. J., Stöcklin, E., Weber, P., Kiely, M., & Hill, T. R. (2012). Relative effectiveness of oral 25-hydroxyvitamin D3 and vitamin D3 in raising wintertime serum 25-hydroxyvitamin D in older adults. The American journal of clinical nutrition, 95(6), 1350-1356.
Hill, T. R., Cotter, A. A., Mitchell, S., Boreham, C. A., Dubitzky, W., Murray, L., ... & Cashman, K. D. (2010). Vitamin D status and parathyroid hormone relationship in adolescents and its association with bone health parameters: analysis of the Northern Ireland Young Heart’s Project. Osteoporosis international, 21(4), 695-700.
Seamans, K. M., Hill, T. R., Wallace, J. M., Horigan, G., Lucey, A. J., Barnes, M. S., ... & Cashman, K. D. (2010). Cholecalciferol supplementation throughout winter does not affect markers of bone turnover in healthy young and elderly adults. The Journal of nutrition, 140(3), 454-460.
| References/Citations: | How the Intact PTH ELISA kit was used: |
| Cholecalciferol Supplementation throughout Winter Does Not Affect Markers of Bone Turnover in Healthy Young and Elderly Adults Kelly M. Seamans, et al., J. Nutr., Mar 2010; 140: 454 - 460. | The Intact PTH ELISA Kit was used to measure the concentration (ng/mL) of parathyroid hormone in adult human serum. |
| Estimation of the dietary requirement for vitamin D in free-living adults 64 y of age Kevin D Cashman et al.,Am. J. Clinical Nutrition, May 2009; 89: 1366 - 1374. | The Intact PTH ELISA Kit was used to measure the concentration (ng/mL) of parathyroid hormone in adult* human serum.*Adults greater than 64 years of age. |
Estimation of the dietary requirement for vitamin D in healthy adults Kevin D Cashman et al.,Am. J. Clinical Nutrition, Dec 2008; 88: 1535 - 1542. | The Intact PTH ELISA Kit was used to measure the concentration (ng/mL) of parathyroid hormone in adult* human serum.*Adults were age 20 to 40 years. |
| Low vitamin D status adversely affects bone health parameters in adolescents Kevin D Cashman et al.,Am. J. Clinical Nutrition, Apr 2008; 87: 1039 - 1044. | The Intact PTH ELISA Kit was used to measure the concentration (ng/mL) of parathyroid hormone in adolescent human serum. |
How It Works
The Intact PTH Immunoassay is a two-site ELISA (Enzyme-Linked Immunosorbent Assay) for the measurement of the biologically intact 84 amino acid chain of PTH. Two different goat polyclonal antibodies to human PTH have been purified by affinity chromatography to be specific for well defined regions on the PTH molecule. One antibody is prepared to bind only the mid-region and C-terminal PTH 39-84 and this antibody is biotinylated. The other antibody is prepared to bind only the N-terminal PTH 1-34 and this antibody is labeled with horseradish peroxidase (HRP) for detection.
Although mid-region and C-terminal fragments are bound by the biotinylated anti-PTH (39-84), only the intact PTH 1-84 forms the sandwich complex necessary for detection. The capacity of the biotinylated antibody and the streptavidin coated microwell both have been adjusted to exhibit negligible interference by inactive fragments, even at very elevated levels. In this assay, calibrators, controls, or patient samples are simultaneously incubated the enzyme labeled antibody and a biotin coupled antibody in a streptavidin-coated microplate well. At the end of the assay incubation, the microwell is washed to remove unbound components and the enzyme bound to the solid phase is incubated with the substrate tetramethylbenzidine (TMB). An acidic stopping solution is then added to stop the reaction and converts the color to yellow. The intensity of the yellow color is directly proportional to the concentration of intact PTH in the sample. A dose response curve of absorbance unit vs. concentration is generated using results obtained from the calibrators. Concentrations of intact PTH present in the controls and patient samples are determined directly from this curve.
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1 mmol/LEDTA(pH 8.0)
因为含有以上两种物质，所以称为TE。
配制分三步：
1)1 M Tris-HCl (pH 8.0) 50 ml的配制：称取Tris碱6.06 g，加超纯水40 ml溶解，滴加浓HCl约2.1 ml调pH至8.0，定容至50 ml。
2)0.5 M EDTA（pH 8.0）50 ml的配制：称取EDTA-Na2·2H2O 9.306 g，加超纯水35 ml，剧烈搅拌，用约1 g NaOH颗粒调pH至8.0，定容至50 ml。（EDTA二钠盐需加入NaOH将pH调至接近8.0时，才会溶解。）
3)1×TE（10 mM Tris-HCl，pH 8.0；1 mM EDTA，pH 8.0）的配制：
作用：
TE缓冲液是弱碱性,对DNA的碱基有保护性,(DNA在它是的稳定性较好,不易破坏其完整性或产生开环及断裂),包括提取好的DNA也要放在TE缓冲液是保存. 10mMTris-Hcl，pH有7.47.68.0三种。
EDTA调到8.0是为了更好溶解，其他只要调到相应pH就可以。Tris在7－8附近缓冲能力很强，所以加8.0的EDTA下去后，不会改变pH。
求采纳为满意回答。
1、 缓冲溶液对反应物的测定没有干扰
2、缓冲组分的浓度为1:1
3、有足够的缓冲容量
4、缓冲溶液的PH应在所需范围内
5、组成缓冲溶液的弱碱PKB和弱酸PKA应接近或等于所需的POH值或PH值（PH+POH=14）
配制
只要知道缓冲对的PH值，和要配制的缓冲液的pH值（及要求的缓冲液总浓度），就能按公式计算[盐]和[酸]的量。这个算法涉及对数换算，较麻烦，前人为减少后人的计算麻烦，已为我们总结出pH值与缓冲液对离子用量的关系并列出了表格。只要我们知道要配制的缓冲液的pH，经查表便可计算出所用缓冲剂的比例和用量。例如配制500nmpH5.8浓度为0.1M磷酸缓冲液。
经查表知pH5.8浓度为0.2M Na2HPO48.0毫升，而0.2M Na2HPO492.0毫升。依此可推论出配制100ml0.1M的磷酸缓冲液需要0.1M Na2HPO48.0毫升，而0.1M Na2HPO4需要92.0毫升。
计算好后，按计算结果准确称好固态化学成分，放于烧杯中，加少量蒸馏水溶解，转移入50ml容量瓶，加蒸馏水至刻度，摇匀，就能得到所需的缓冲液。
各种缓冲溶液的配制，均按表格按比例混合，某些试剂，必须标定配成准确浓度才能进行，如醋酸、氢氧化钠等。另外，所有缓冲溶剂的配制计量都能从以上的算式准确获得。
