S100鈣結(jié)合蛋白A12(S100A12)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法)
ELISA Kit for S100 Calcium Binding Protein A12 (S100A12)
CGRP; P6; CAAF1; CAGC; ENRAGE; MRP6; Calgranulin C; Calcium-binding protein in amniotic fluid 1; Extracellular newly identified RAGE-binding; Migration inhibitory factor-related 6
- 編號(hào)SEB080Hu
 - 物種Homo sapiens (Human,人)相同的名稱,不同的物種。
 - 實(shí)驗(yàn)方法雙抗夾心
 - 反應(yīng)時(shí)長(zhǎng)3h
 - 檢測(cè)范圍78-5,000pg/mL
 - 靈敏度最小可檢測(cè)劑量小于等于33pg/mL.
 - 樣本類型serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
 - 下載英文說明書 中文說明書
 - 規(guī)格48T96T96T*596T*1096T*100
 - 價(jià)格¥ 2352¥ 3360¥ 15120¥ 28560¥ 235200
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特異性
                        本試劑盒用于檢測(cè)S100鈣結(jié)合蛋白A12(S100A12),經(jīng)檢測(cè)與其它相似物質(zhì)無明顯交叉反應(yīng)。
                        由于受到技術(shù)及樣本來源的限制,不可能完成對(duì)所有相關(guān)或相似物質(zhì)交叉反應(yīng)檢測(cè),因此本試劑盒有可能與未經(jīng)檢測(cè)的其它物質(zhì)有交叉反應(yīng)。
                    
回收率
分別于定值血清及血漿樣本中加入一定量的S100鈣結(jié)合蛋白A12(S100A12)(加標(biāo)樣品),重復(fù)測(cè)定并計(jì)算其均值,回收率為測(cè)定值與理論值的比率。
| 樣本 | 回收率范圍(%) | 平均回收率(%) | 
| serum(n=5) | 98-105 | 102 | 
| EDTA plasma(n=5) | 81-104 | 101 | 
| heparin plasma(n=5) | 79-101 | 82 | 
精密度
                    精密度用樣品測(cè)定值的變異系數(shù)CV表示。CV(%) = SD/mean×100 
                    批內(nèi)差:取同批次試劑盒對(duì)低、中、高值定值樣本進(jìn)行定量檢測(cè),每份樣本連續(xù)測(cè)定20 次,分別計(jì)算不同濃度樣本的平均值及SD值。
                    批間差:選取3個(gè)不同批次的試劑盒分別對(duì)低、中、高值定值樣本進(jìn)行定量測(cè)定,每個(gè)樣本使用同一試劑盒重復(fù)測(cè)定8次,分別計(jì)算不同濃度樣本的平均值及SD值。
                    批內(nèi)差: CV<10% 
                    批間差: CV<12% 
線性
在定值血清及血漿樣本內(nèi)加入適量的S100鈣結(jié)合蛋白A12(S100A12),并倍比稀釋成1:2,1:4,1:8,1:16的待測(cè)樣本,線性范圍即為稀釋后樣本中S100鈣結(jié)合蛋白A12(S100A12)含量的測(cè)定值與理論值的比率。
| 樣本 | 1:2 | 1:4 | 1:8 | 1:16 | 
| serum(n=5) | 87-101% | 89-101% | 79-101% | 90-97% | 
| EDTA plasma(n=5) | 80-104% | 92-102% | 78-98% | 86-95% | 
| heparin plasma(n=5) | 83-99% | 88-102% | 85-103% | 93-101% | 
穩(wěn)定性
                    經(jīng)測(cè)定,試劑盒在有效期內(nèi)按推薦溫度保存,其活性降低率小于5%。
                    為減小外部因素對(duì)試劑盒破壞前后檢測(cè)值的影響,實(shí)驗(yàn)室的環(huán)境條件需盡量保持一致,尤其是實(shí)驗(yàn)室內(nèi)溫度、濕度及溫育條件。其次由同一實(shí)驗(yàn)員來進(jìn)行操作可減少人為誤差。
                
實(shí)驗(yàn)流程
1. 實(shí)驗(yàn)前標(biāo)準(zhǔn)品、試劑及樣本的準(zhǔn)備;
                            2. 加樣(標(biāo)準(zhǔn)品及樣本)100µL,37°C孵育1小時(shí);
                            3. 吸棄,加檢測(cè)溶液A100µL,37°C孵育1小時(shí);
                            4. 洗板3次;
                            5. 加檢測(cè)溶液B100µL,37°C孵育30分鐘;
                            6. 洗板5次;
                            7. 加TMB底物90µL,37°C孵育10-20分鐘;
                            8. 加終止液50µL,立即450nm讀數(shù)。
實(shí)驗(yàn)原理
將S100鈣結(jié)合蛋白A12(S100A12)抗體包被于96孔微孔板中,制成固相載體,向微孔中分別加入標(biāo)準(zhǔn)品或標(biāo)本,其中的S100鈣結(jié)合蛋白A12(S100A12)與連接于固相載體上的抗體結(jié)合,然后加入生物素化的S100鈣結(jié)合蛋白A12(S100A12)抗體,將未結(jié)合的生物素化抗體洗凈后,加入HRP標(biāo)記的親和素,再次徹底洗滌后加入TMB底物顯色。TMB在過氧化物酶的催化下轉(zhuǎn)化成藍(lán)色,并在酸的作用下轉(zhuǎn)化成最終的黃色。顏色的深淺和樣品中的S100鈣結(jié)合蛋白A12(S100A12)呈正相關(guān)。用酶標(biāo)儀在450nm波長(zhǎng)下測(cè)定吸光度(O.D.值),計(jì)算樣品濃度。
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參考文獻(xiàn)
| 雜志 | 參考文獻(xiàn) | 
| Inflammation Research | Correlation of human S100A12 (EN-RAGE) and high-sensitivity C-reactive protein as gingival crevicular fluid and serum markers of inflammation in chronic periodontitis and type 2 diabetes[Pubmed: 24378957] | 
| Dig Dis Sci | The Circulating Level of Soluble Receptor for Advanced Glycation End Products Displays Different Patterns in Ulcerative Colitis and Crohn’s Disease: A Cross-Sectional Study[PubMed: 25757448] | 
| The Journal of Clinical Endocrinology & Metabolism | Low Endogenous Secretory Receptor for Advanced Glycation End-Products Levels Are Associated With Inflammation and Carotid Atherosclerosis in Prediabetes[Pubmed:26885882] | 
| Nutrition, Metabolism, and Cardiovascular Diseases | High intake of dietary advanced glycation end-products is associated with increased arterial stiffness and inflammation in subjects with type 2 diabetes.[pubmed:28958695] | 
| The Journal of Clinical Endocrinology & Metabolism | HbA1c Identifies Subjects With Prediabetes and Subclinical Left Ventricular Diastolic Dysfunction.[pubmed:28973588] | 
| Journal of Immunology Research | A Granulocyte-Specific Protein S100A12 as a Potential Prognostic Factor Affecting Aggressiveness of Therapy in Patients with Juvenile Idiopathic Arthritis[Doi: 10.1155/2018/5349837] | 
| Al-Azhar Assiut Medical Journal | The role of serum S100A12 (calgranulin C) as a diagnostic marker in Egyptian patients with irritable bowel syndrome and ulcerative colitis[Doi: 10.4103/AZMJ.AZMJ_52_18] | 
| Acta?Diabetologica | Analysis of S100A12 plasma levels in hyperlipidemic subjects with or without familial hypercholesterolemia[Pubmed: 30963307] | 
| Cells | 1 h Postload Glycemia Is Associated with Low Endogenous Secretory Receptor for Advanced Glycation End Product Levels and Early Markers of Cardiovascular?…[] | 
| Biomolecules . | Multimarker Approach to Identify Patients with Coronary Artery Disease at High Risk for Subsequent Cardiac Adverse Events: The Multi-Biomarker Study[Pubmed: 32549327] | 
| Mediators of Inflammation | Alarmins HMGB1, IL-33, S100A7, and S100A12 in Psoriasis Vulgaris[] | 
| Animals (Basel) | The Serum and Saliva Proteome of Dogs with Diabetes Mellitus[33271797] | 





