| Kitsize | 12x8 |
| Method | ELISA |
| Incubationtime | 1x2h,1x30min |
| Standardrange | 1.7-420ng/mL |
| Specimen/Volumes | 10µLurine |
| Substrate/isotope | TMB450nm |
| RegulatoryStatus: | EU:CE,CDN:IVD |
Melatonin–Morethanasleephormone
IBLInternationalistheonlycommercialsupplierofacompletesetofuserfriendlyimmunoassaysforthereliablequantitativeassessmentofmelatonininserumaswellasinsaliva,andofitsprimarymetabolite6-sulfatoxymelatonininurine.
- TheassaysarecalibratedtoLC-MS/MS
- Thebroadstandardrangeallowsapplicationfortherapeuticdrugmonitoring
- Validatedreferencevaluesforurinary6-sulfatoxymelatonin(figure1)1
- Successfullyappliedindietresearch2-4,variousspeciesandcellculturesupernatants*
- Widelyusedandcitedinmultiplepublications*

Timeandagedependentreferencevaluesofurinary6-sulfatoxymelatoninmeasuredwithIBLInternational’sMelatonin-SulfateUrineELISA1
Melatonin:aubiquitouslydistributedandfunctionallydiversemolecule
Melatoninisanendogenoushormonesynthesizedandsecretedprimarilybythepinealgland.Itssecretionisregulatedbythesuprachiasmaticnucleiandregulatesthediurnallight-darkcyclewithpeakproductionatnightandlowlevelduringthedaytime.
Apartfromitsroleintheregulationofsleep,multiplestudiesshowitsinvolvementinawidevarietyofphysiologicalprocesses(affectingcircADIanrhythmandmood,interactingwiththeimmunesystemandactingasantioxidant,freeradicalscavengerandoncostaticagent).Moreover,ithasbeenshownthatmelatoninperformsthesefunctionsbyavarietyofmechanisms.
- MahlbergR.etal.Normativedataonthedailyprofileofurinary6-sulfatoxymelatonininhealthysubjectsbetweentheagesof20and84.Psychoneuroendocrinology.2006Jun;31(5):634-41.
- DelaPuertaC.etal.Melatoninisaphytochemicalinoliveoil.FoodChem.2007;104:609-12.
- ChoMH.etal.Chitosantreatmentsaffectgrowthandselectedqualityofsunflowersprouts.JFoodSci.2008Jan;73(1):S70-7.
- MaldonadoMD.etal.Melatoninpresentinbeercontributestoincreasethelevelsofmelatoninandantioxidantcapacityofthehumanserum.ClinNutr.2009Apr;28(2):188-91.
*ThepublicationscanberequestedatIBLinternationalviadan@ibl-international.com
Enzymeimmunoassayforthein-vitrodiagnosticquantitativedeterminationofMelatoninSulfate(synonyms:6-hydroxymelatoninsulfate,6-sulfatoxymelatonin)inhumanurine.Thepinealgland(corpuspineale)hasbeencalledaneuroendocrinetransducerbecauseofitsimportantroleinphotoperiodism.ThemajorhormoneofthepinealglandisN-acetyl-5-methoxy-tryptamineormelatoninwhichissynthezisedfromtheaminoacidtryptophane.Melatoninhasitshighestlevelsinplasmaduringnighttime.Itscharacteristicnocturnalsurgeappearstoencodetemporalinformationsuchaslengthofnight.Regulationofthemelatoninsecretionisunderneuralcontrol.Sympatheticinnervationseemstoplayamajorroleviaitsreleaseofnoradrenaline.Alteredpatternsand/orlevelsofmelatoninsecretionhavebeenreportedtocoincidewithsleepdisorders,jetlag,depression,stress,schizophrenia,hypothalamicamenorrhea,pregnancy,anorexianervosa,someformsofcancer,immunologicaldisordersaswellascontrolofsexualmaturationduringpuberty.Mostofthecirculatingmelatoninismetabolizedintheliverto6-hydroxymelatoninandsubsequentlyto6-sulfatoxymelatoninwhichisexcretedintotheurine.Theconcentrationof6-hydroxymelatoninsulfateinurinecorrelateswellwiththetotallevelofmelatonininthebloodduringthecollectionperiod.
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1. 直接法测定抗原 2.间接法测定抗体 3.双抗体夹心法测定抗原4. 竞争法测定抗原 厚百生物专业提供各类生化实验试剂、仪器、耗材,提供技术服务,满足您的实验室常规采购需求。厚百,让您更省心!
(1) 包被:用0.05M PH9.牰碳酸盐包被缓冲液将抗体稀释至蛋白质含量为1~10μg/ml。在每个聚苯乙烯板的反应孔中加0.1ml,4℃过夜。次日,弃去孔内溶液,用洗涤缓冲液洗3次,每次3分钟。(简称洗涤,下同)。
(2) 加样:加一定稀释的待检样品0.1ml于上述已包被之反应孔中,置37℃孵育1小时。然后洗涤。(同时做空白孔,阴性对照孔及阳性对照孔)。
(3) 加酶标抗体:于各反应孔中,加入新鲜稀释的酶标抗体(经滴定后的稀释度)0.1ml。37℃孵育0.5~1小时,洗涤。
(4) 加底物液显色:于各反应孔中加入临时配制的TMB底物溶液0.1ml,37℃10~30分钟。
(5) 终止反应:于各反应孔中加入2M硫酸0.05ml。
(6) 结果判定:可于白色背景上,直接用肉眼观察结果:反应孔内颜色越深,阳性程度越强,阴性反应为无色或极浅,依据所呈颜色的深浅,以“+”、“-”号表示。也可测O·D值:在ELISA检测仪上,于450nm(若以ABTS显色,则410nm)处,以空白对照孔调零后测各孔O·D值,若大于规定的阴性对照OD值的2.1倍,即为阳性。
间接法:
用包被缓冲液将已知抗原稀释至1~10μg/ml,每孔加0.1ml,4℃过夜。次日洗涤3次。加一定稀释的待检样品(未知抗体)0.1ml于上述已包被之反应孔中,置37℃孵育1小时,洗涤。(同时做空白、阴性及阳性孔对照)于反应孔中,加入新鲜稀释的酶标第二抗体(抗抗体)0.1ml,37℃孵育30-60分钟,洗涤,最后一遍用DDW洗涤。其余步骤同“双抗体夹心法”的4、5、6。
表面抗体是双抗原夹心法
E抗体和核心抗体是竞争抑制法或间接法
1. 包被:用0.05M PH9,碳酸盐包被缓冲液将抗体稀释至蛋白质含量为1~10μg/ml,在每个聚苯乙烯板的反应孔中加0.1ml,4℃过夜。次日,弃去孔内溶液,用洗涤缓冲液洗3次,每次3分钟。
2. 加样:加一定稀释的待检样品0.1ml于上述已包被之反应孔中,置37℃孵育1小时。然后洗涤。(同时做空白孔,阴性对照孔及阳性对照孔)。
3. 加酶标抗体:于各反应孔中,加入新鲜稀释的酶标抗体(经滴定后的稀释度)0.1ml,37℃孵育0.5~1小时,洗涤。
4. 加底物液显色:于各反应孔中加入临时配制的TMB底物溶液0.1ml,37℃10~30分钟。
5. 终止反应:于各反应孔中加入2M硫酸0.05ml。
6. 结果判定:可于白色背景上,直接用肉眼观察结果:反应孔内颜色越深,阳性程度越强,阴性反应为无色或极浅,依据所呈颜色的深浅,以“+”、“-”号表示。也可测OD值:在ELISA检测仪上,于450nm(若以ABTS显色,则410nm)处,以空白对照孔调零后测各孔OD值,若大于规定的阴性对照OD值的2.1倍,即为阳性。
方法二:用于检测未知抗体的间接法:
用包被缓冲液将已知抗原稀释至1~10μg/ml,每孔加0.1ml,4℃过夜。次日洗涤3次。加一定稀释的待检样品(未知抗体)0.1ml于上述已包被之反应孔中,置37℃孵育1小时,洗涤。(同时做空白、阴性及阳性孔对照)于反应孔中,加入新鲜稀释的酶标第二抗体(抗抗体)0.1ml,37℃孵育30~60分钟,洗涤,最后一遍用DDW洗涤。其余步骤同“双抗体夹心法”的4、5、6。

