粘蛋白5B(MUC5B)检测试剂盒(酶联免疫吸附试验法)

ELISA Kit for Mucin 5 Subtype B (MUC5B)

MUC5-B; MUC9; MG1; Oligomeric Mucus/Gel-Forming; Mucin 5,Subtype B,Tracheobronchial; Cervical mucin; High molecular weight salivary mucin MG1; Sublingual gland mucin

特异性

本试剂盒用于检测粘蛋白5B(MUC5B),经检测与其它相似物质无明显交叉反应。
由于受到技术及样本来源的限制,不可能完成对所有相关或相似物质交叉反应检测,因此本试剂盒有可能与未经检测的其它物质有交叉反应。

精密度

精密度用样品测定值的变异系数CV表示。CV(%) = SD/mean×100
批内差:取同批次试剂盒对低、中、高值定值样本进行定量检测,每份样本连续测定20 次,分别计算不同浓度样本的平均值及SD值。
批间差:选取3个不同批次的试剂盒分别对低、中、高值定值样本进行定量测定,每个样本使用同一试剂盒重复测定8次,分别计算不同浓度样本的平均值及SD值。
批内差: CV<10%
批间差: CV<12%

稳定性

经测定,试剂盒在有效期内按推荐温度保存,其活性降低率小于5%。
为减小外部因素对试剂盒破坏前后检测值的影响,实验室的环境条件需尽量保持一致,尤其是实验室内温度、湿度及温育条件。其次由同一实验员来进行操作可减少人为误差。

实验流程

1. 实验前标准品、试剂及样本的准备;
2. 加样(标准品及样本)100µL,37°C孵育1小时;
3. 吸弃,加检测溶液A100µL,37°C孵育1小时;
4. 洗板3次;
5. 加检测溶液B100µL,37°C孵育30分钟;
6. 洗板5次;
7. 加TMB底物90µL,37°C孵育10-20分钟;
8. 加终止液50µL,立即450nm读数。

实验原理

将粘蛋白5B(MUC5B)抗体包被于96孔微孔板中,制成固相载体,向微孔中分别加入标准品或标本,其中的粘蛋白5B(MUC5B)与连接于固相载体上的抗体结合,然后加入生物素化的粘蛋白5B(MUC5B)抗体,将未结合的生物素化抗体洗净后,加入HRP标记的亲和素,再次彻底洗涤后加入TMB底物显色。TMB在过氧化物酶的催化下转化成蓝色,并在酸的作用下转化成最终的黄色。颜色的深浅和样品中的粘蛋白5B(MUC5B)呈正相关。用酶标仪在450nm波长下测定吸光度(O.D.值),计算样品浓度。

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参考文献

杂志参考文献
The Journal of Biological ChemistryThe Extracellular Signal-regulated Kinase Mitogen-activated Protein Kinase/Ribosomal S6 Protein Kinase 1 Cascade Phosphorylates cAMP Response Element-binding Protein to Induce MUC5B Gene Expression via D-Prostanoid Receptor Signaling[Jcb: 34199]
The Journal of Biological ChemistryThe ERK MAPK/RSK1 cascade phosphorylates CREB to induce MUC5B gene expression via DP1 receptor signaling[Jcb: 247684]
Respiratory ResearhPyocyanin-induced mucin production is associated with redox modification of FOXA2[PubMed: PMC3765780]
RespirologySecreted mucins and airway bacterial colonization in non-CF bronchiectasis[PubMed: 26172851]
American Thoracic Society International Conference AbstractsMucin Secretion In Asthmatics With Osmotic Airway Hyperresponsiveness[doi:10.1164]
Ann Am Thorac SocAirway Mucin 2 Is Decreased in Patients with Severe Chronic Obstructive Pulmonary Disease with Bacterial Colonization[doi:10.1513]
Секреторно-экссудативная реакция слизистой носа на воздействие холодного воздуха у больных бронхиальной астмой[10.20538/1682-0363-2017-2-146-158]
AllergyTh2 cytokines orchestrate the secretion of MUC 5 AC and MUC 5B in IL‐5 positive chronic rhinosinusitis with nasal polyps[Pubmed:29802623]
European Journal of Pharmacology Salvianolic acid B improves airway hyperresponsiveness by inhibiting MUC5AC overproduction associated with Erk1/2/P38 signaling[Pubmed:29382534]
Japan Society of Immunology & Allergology in Otolaryngologyインフルエンザ菌Phosphorylcholine の表出と中耳ムチンMUC5AC 及びMUC5B 産生への影響[10.5648:jjiao.36.7]
Journal of Investigative MedicineThe characteristics and clinical significance of mucin levels in bronchoalveolar lavage fluid of patients with interstitial lung disease[Pubmed: 30573494]
Mucosal ImmunologyASC and NLRP3 maintain innate immune homeostasis in the airway through an inflammasome-independent mechanism[Pubmed: 31278375]