- Overview
- Data/Specifications
- Literature/Support
- How It Works
- Related Products
Overview
β-endorphin is an endogenous opioid peptide neurotransmitter found in the neurons of both the central and peripheral nervous system. It is a 31 amino acids long resulting from processing of the precursor proopiomelanocortin (POMC), which also gives rise to the other peptide hormones including ACTH (adrenocorticotropic hormone) andα- andγ-MSH (melanocyte-stimulating hormone).β-endorphin is an agonist of opioid receptors giving it analgesic properties.
Data/Specifications
Species: human
Sample Type:serum, plasma
Sample Preparation:plasma extraction
Sample Size:50 uL
Standard Curve Range: 0.01 - 100 ng/mL
Sensitivity: 1 ng/mL
Assay Length: 4 hrs
Cross-reactivity:
Leu-Enkephalin: 0 %
MET-Enkephalin: 0 %
beta_Endorphin (Human): 100 %
alpha Endorphin: 0 %
Gamma Endorphin: 0 %
Literature/Support
Product Insert:
β-endorphin ELISA Insert (PDF)
Articles/Troublshooting:
ELISA Troubleshooting Guide
ELISA Data Reduction Guide
Appel, C. K., Scheff, N. N., Viet, C. T., Schmidt, B. L., & Heegaard, A. M. (2019).Decitabine attenuates nociceptive behavior in a murine model of bone cancer pain.Pain,160(3), 619-631.
Setiawardhani, A. L., Srilestari, A., & Simadibrata, C. (2017, August). Electroacupuncture effect at the LI 4 Hegu point on the plasmaβ-endorphin level of healthy subjects. InJournal of Physics: Conference Series(Vol. 884, No. 1, p. 012027). IOP Publishing.
Viet, C. T., Dang, D., Ye, Y., Ono, K., Campbell, R. R., & Schmidt, B. L. (2014). Demethylating drugs as novel analgesics for cancer pain. Clinical Cancer Research, 20(18), 4882-4893.
Milman, S., Leu, J., Shamoon, H., Vele, S., & Gabriely, I. (2012). Opioid receptor blockade prevents exercise-associated autonomic failure in humans. Diabetes, 61(6), 1609-1615.
Milman, S., Leu, J., Shamoon, H., Vele, S., & Gabriely, I. (2011). Magnitude of exercise-induced β-endorphin response is associated with subsequent development of altered hypoglycemia counterregulation. The Journal of Clinical Endocrinology & Metabolism, 97(2), 623-631.
Vele, S., Milman, S., Shamoon, H., & Gabriely, I. (2011). Opioid receptor blockade improves hypoglycemia-associated autonomic failure in type 1 diabetes mellitus. The Journal of Clinical Endocrinology & Metabolism, 96(11), 3424-3431.
Viet, C. T., Ye, Y., Dang, D., Lam, D. K., Achdjian, S., Zhang, J., & Schmidt, B. L. (2011). Re-expression of the methylated EDNRB gene in oral squamous cell carcinoma attenuates cancer-induced pain. PAIN®, 152(10), 2323-2332.
| References/Citations: | The Beta-Endorphin ELISA Kit was used for: |
| Validation of a sham acupuncture procedure in a randomised, controlled clinical trial of chronic pelvic pain treatmentLee, SW et al., Acupunct Med. 2011 | Measure the concentration of beta-endorphin in accupuncture treated patient group. |
| Vernonia cinerea Less. supplementation and strenuous exercise reduce smoking rate: relation to oxidative stress status and beta-endorphin release in active smokers. Leelarungrayub D, et al. J Int Soc Sports Nutr. 2010 May; 7:21. | Measure the concentration of beta-endorphin in human plasma samples. |
| Propofol/alfentanil and propofol/ketamine procedural sedation in children with acute lymphoblastic leukaemia: safety, efficacy and their correlation with pain neuromediator expression. Chiaretti A, et al. Eur J Cancer Care. 2010 Mar; 19(2):212-20. | |
Endothelin-A Receptor Antagonism Attenuates Carcinoma-Induced Pain Through Opioids in Mice. Quang PN and Schmidt BL. J Pain. 2010 Jan (Epub ahead of print) | Measure the concentration of BE in media collected from squamous cell carcinoma (SCC) cell culture.The SCC cell line was derived from human tongue. |
| The Effects of Hydrotherapy on Anxiety, Pain, Neuroendocrine Responses, and Contraction Dynamics During Labor. Benfield RD, et al. Biol Res Nurs. 2010 May | Measure the concentration of BE in plasma samples from female humans.Plasma was prepared from whole blood collected in vacutainers containing EDTA.Traysylol (5,000 KIU) was added to inhibit proteolysis.Blood was spun at 3,000 rpm for 15 minutes at 14 °C and the plasma was collected, stored at -80 °C. |
| Biologically active breast milk proteins in association with very preterm delivery and stage of lactation. Mehta R and Petrova A. J Perinatol. 2010 Jun | Measure the concentration of beta-endorphin in breast milk samples obtained from female humans. |
| Serum beta-endorphin response to stress before and after operation under fentanyl anesthesia in neonates, infants and preschool children. Mirilas P, et al. Eur J Pediatr Surg. 2010 Mar; 20(2):106-10. | Measure concentration of serum BE in human samples. |
| Clinical and endocrinological changes after electro-acupuncture treatment in patients with osteoarthritis of the knee. Ahsin S, et al. Pain. 2009 Dec; 147(1-3):60-6. | Measure the concentration of beta-endorphin in plasma from human patients with osteoarthritis of the knee.Plasma was obtained from blood collected in a pre-cooled EDTA tube. |
| Peripheral endothelin B receptor agonist-induced antinociception involves endogenous opioids in mice. Quang PN and Schmidt BL. Pain. 2010 May; 149(2):254-62. | Measure the concentration of BE in media collected from squamous cell carcinoma (SCC) cell culture.The SCC cell line was derived from human tongue. |
| Beta-endorphin in serum and seminal plasma in infertile men. El-Hagger S, et al. Asian J Androl. 2006 Jun; 8(6):709-12. | Measure the concentration of beta-endorphin in seminal plasma and serum from male human patients. |
| Cognitive and emotional factors in placebo analgesia. Flaten MA, et al. J Psychosom Res. 2006 Jul; 61(1):81-9. | Measure the concentration of serum BE from adult humans.Serum was prepared from whole blood by centrifugation at 1500g for 10 minutes. |
Plasma beta-endorphin levels in frequent and infrequent tanners before and after ultraviolet and non-ultraviolet stimuli. Kaur M, et al. J Am Acad Dermatol. 2006 May;54(5):919-20. | Measure the concentration of beta-endorphin in plasma from female human subjects exposed to Ultraviolet light. |
| Treatment with double dose of omezprazole increases Beta Endorphin plasma in patients with coronary artery disease. Budzynski J, et al. Archives of Medical Science. 2010; 6(2):201-207. |
How It Works
The immunoplate in this kit is pre-coated with secondary antibody and the nonspecific binding sites are blocked. The secondary antibody can bind to the Fc fragment of the primary antibody (peptide antibody) whose Fab fragment will be competitively bound by both biotinylated peptide and peptide standard or targeted peptide in sample. The biotinylated peptide is able to interact with streptavidin-horseradish peroxidase (SA-HRP) which catalyzes the substrate solution composed of 3,3’,5,5’-tetramethylbenzidine (TMB) and hydrogen peroxide to produce a blue colored solution. The enzyme-substrate reaction is stopped by hydrogen chloride (HCl) and the solution turns to yellow. The intensity of the yellow is directly proportional to the amount of biotinylated peptide-SA/HRP complex but inversely proportional to the amount of the peptide in standard solutions or samples. This is due to the competitive binding of the biotinylated peptide and the peptide in standard solutions or samples to the peptide antibody (primary antibody). A standard curve of a peptide with known concentration can be established accordingly. The peptide with unknown concentration in samples can be determined by extrapolation to this standard curve.
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实时定量的引物设计要求比较高，可以按普通引物设计的方法来做，但是设计时要注意扩增的片段不能太大，最好在250bp-100bp之间。同时要保证这对引物有绝对的特异性，因为要是产生非特异性扩增的话，就不能准确的计量模板量了，这样就失去了实时定量的意义了
用已知浓度的DNA样本（通常是质粒）梯度稀释成一系列的样本。做实验的时候，这些标准品和待测样本加在同样一块96孔盘的不同孔内。一起做PCR。
标准品的荧光强度和已知的浓度作图，可以得到一条标准曲线。而待测样本的荧光强度测出以后，就可以在标准曲线上算出样本浓度了。
目前各家仪器公司推出自己的数字PCR，目前市场主流的是三家，1、LIFETechnologies3D数字PCR（芯片式数字PCR）：我个人认为是微孔数字PCR，主要是将20ul反应体系分散到20000个微孔中进行反应,变成20000个反应体系，PCR反应结束后，采用CCD拍照，数阳性反应孔。2、Bio-Rad的微滴数字PCR（油包水原理）：将20ul反应体系采用液滴反应器形成20000个油包水反应体系，PCR反应结束后，才有流式细胞术的原理，检测每一个液体，数阳性反应的液滴数量。3、RainDance的数字PCR（油包水原理）、原理与伯乐微滴数字PCR相同，但是其液体形成能力比伯乐强很多，理论上可以产生1000万个小油滴。价格从高到底：Raindance、Bio-Rad、LIFETechnologies。目前本人使用的是Bio-RadQX200、正在做实验室，希望广数字PCR的使用者可以相互交流，相互解答疑问，更好的利用数字PCR。
为适应医疗器械监督管理工作的需要，总局组织有关单位和专家对恒温核酸扩增检测仪等22个产品的管理类别进行了界定。现通知如下：
一、作为Ⅲ类医疗器械管理的产品（1个）
恒温核酸扩增检测仪：由检测系统、加热模块、温控系统、触摸屏和随机软件组成。基于荧光检测的恒温核酸扩增检测技术，定性检测样本中的目标核酸有无扩增。分类编码：6840。
二、作为Ⅱ类医疗器械管理的产品（7个）
（一）免疫磁微粒捕获仪：由蠕动泵、管路、永磁铁及其他必要辅助器具组成。用于磁微粒免疫分析中捕获磁微粒复合物，是临床免疫磁微粒检验分析的前处理设备。分类编码：6841。
（二）精液采集器：由精液收集套、润滑剂、精液采集管、采集漏斗、袖珍剪组成。产品以无菌型式提供。临床上用于收集成年男性精液，为评价男性生育能力提供完整标本。分类编码：6841。
（三）干式血细胞微量样品测试管：由测试管体、封管器、浮子、肝素钠抗凝剂和荧光吖啶橙染料组成。用于收集末梢血样和静脉血样，临床上与干式血细胞计数仪配合使用，对样本染色、抗凝和分类。分类编码：6841。
（四）胎盘生长因子检测试剂盒（时间分辨荧光法）：由校准品、铕标示踪抗体、分析缓冲液、生物素抗体、链霉亲和素微孔板条、条形码标签等组成。用于定量测定孕妇血清中的胎盘生长因子。临床上用于辅助筛查妊娠妇女孕早期唐氏综合症风险。分类编码：6840。
（五）脂联素检测试剂盒（免疫比浊法）：由试剂1（缓冲液）和试剂2（乳胶抗体试剂）组成。用于体外定量检测人血清或血浆中脂联素浓度。临床上用于评估2型糖尿病和心血管疾病的风险。分类编码：6840。
（六）小而密低密度脂蛋白胆固醇测定试剂盒（直接清除法）：由试剂（含过氧化氢酶、胆固醇氧化酶、胆固醇脂酶、显色剂的缓冲液）和试剂（含过氧化物酶、酶稳定剂、显色剂的缓冲液）组成。用于体外定量检测人血清中小而密低密度脂蛋白胆固醇的含量，临床上用于动脉粥样硬化的辅助诊断。分类编码：6840。
（七）尿液游离LamBDa-轻链测定试剂盒（免疫比浊法）：由定标液、试剂缓冲液和试剂（游离Lambda-轻链抗血清）组成。用于体外定量测定尿液游离Lambda-轻链浓度，临床上用于单克隆丙种球蛋白增多症的辅助诊断。分类编码：6840。
三、作为第Ⅰ类医疗器械管理的产品（13个）
（一）制片染色一体机：通常由离心模块、制片模块、染色模块、控制系统等功能模块组成。用于病理分析前对人体细胞或细菌样本进行制片及染色。分类编码：6841。
（二）细胞分离制片染色一体机：通常由控制系统、过滤器、多路阀注射泵、贮液瓶、废液瓶等组成。适用于病理分析前对人体外周血或组织样本中细胞的分离、过滤、制片和染色。分类编码：6841。
（三）试剂卡孵育器：由塑料外壳、温控模块、电器元件组成。用于特定试剂卡的孵育。分类编码：6840。
（四）细胞过滤采集器：由软管、过滤收集病变细胞装置、抽吸装置组成。用于病理分析前对人体内脱落病变细胞的采集。分类编码：6841。
（五）液基细胞和微生物处理、保存试剂：主要由细胞保存液、细胞裂解液、提取液、稀释液、消化酶、防腐剂等及必要的细胞承载或制片器具组成。用于临床检验分析前细胞或微生物的保存、运输、提取、分离、沉淀、固定、制片等。分类编码：6840。
（六）病理分析前处理试剂：主要由组织固定液、组织脱水液、透明液、清洗剂等组成。用于病理分析前组织标本的固定、梯度脱水、透明、浸蜡和包埋制片。分类编码：6840。
（七）脱蜡液：由脱蜡液、防腐液、专用水等组成。用于对样本进行染色前预处理，去除石蜡包埋组织样本上的石蜡。分类编码：6840。
（八）触发缓冲液：由0.9％的氯化钠等渗溶液组成。与血小板功能分析仪及配套检测试剂盒配套使用，用于在检测前润湿生化活性膜。分类编码：6840。
（九）流式细胞分析用鞘液：由含盐缓冲液和防腐剂组成。用于与流式技术相关的分析中对样本进行检测时形成鞘流，以利于仪器进行计数分析。分类编码：6840。
（十）流式细胞分析用溶血剂：由浓度小于2％的甲醛和浓度小于15％的甘油组成。用于快速裂解人类外周血中红细胞，维持白细胞基本形态，临床上用于流式细胞仪检测前样本处理。分类编码：6840。
（十一）抗体稀释液：由缓冲液和防腐剂组成。用于临床检验时抗血清的稀释。分类编码：6840。
（十二）溶痰剂：由次氯酸钠、表面活性剂组成。用于抗酸染色前痰标本的均质化处理，以提高阳性检出率。分类编码：6840。
（十三）精液液化剂：由菠萝蛋白酶和蔗糖组成。用于精液检查前促进液化迟缓的精液标本的液化、降低精液的粘稠度。分类编码：6840。
四、不按照医疗器械管理的产品（1个）
瓷珠菌种保存管：由表面多孔的小瓷珠、胰蛋白脉大豆肉汤、甘油和蔗糖按一定比例混合组成。用于微生物检验中菌种的保存。
自本文件发布之日起，对于不作为医疗器械管理的，如已受理尚未完成注册审批的，食品药品监管部门应按规定不予注册，相关注册申请资料予以存档。尚在有效期内的医疗器械注册证书不得继续使用。
食品药品监管总局办公厅
2015年6月11日
还需要反转录用的引物、定量PCR用的一对引物、荧光信号源（荧光染料或荧光标记的探针，看你用什么策略来做定量PCR）
报价要看你需要服务商提供什么服务，如果你只提供实验材料，要求服务商做以上全部工作，只检测一个指标（一个基因的表达水平变化），价格大约每个样品350-500元，多一个指标大约多90元。
一般至少做一组数据两个样品嘛，所以你按1000块一组样品预算吧。
