
Recombinant Human HA-GATE-16 Vinyl Sulfone Protein, CF Summary
Product Datasheets
Carrier Free
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins.Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration.The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard.In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
UL-446
Shipping | The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. |
Stability & Storage: | Store the unopened product at -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date. |
Reconstitution Calculator
Background: GATE-16
Golgi-associated ATPase Enhancer of 16 kDa (GATE-16), also known as Apg8p2 and GABARAPL2, is a 117 amino acid (aa) polypeptide and a member of the Autophagy-related 8 (Atg8) family of proteins (1). GATE-16/Apg8p2 has 100% aa sequence identity with its mouse and rat orthologs, and is orthologous to the yeast Atg8. Atg8 family members show structural similarity with Ubiquitin, but lack aa sequence similarity. GATE-16/Apg8p2 is best known for its role in autophagy (2,3). GATE-16/Apg8p2 covalently attaches to phosphatidylethanolamine (PE) the phagophore (autophagosome precursor) membrane using a Ubiquitin-like conjugation system that includes Ubiquitin-activating (E1)-, Ubiquitin-conjugating (E2)-, and Ubiquitin Ligase (E3)-like enzymes. Here it is involved in the later stages of autophagosome formation (4,5). It may also be involved in cargo recruitment to autophagosomes (1).
This N-terminal HA-tagged Apg8 protein is a potent, irreversible and specific inhibitor of Agp8-specific isopeptidases (such as Apg84B, Catalog #E-400). Apg84B activities include the processing of Apg8 precursor proteins and the removal of Apg8 proteins that are conjugated to phosphatidylethanolamine during autophagy. These processes can be inhibited by this vinyl sulfone derivative which reacts with the Apg84B active site cysteine. The HA peptide sequence (YPYDVPDYA) is derived from the influenza hemagglutin in protein. This epitope allows for the sensitive identification or purification of such deconjugating activities since it is specifically recognized by anti-HA antibodies and/or anti-HA-agarose.
FAQs
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优点:
1、反应杯自动加载;
2、样本载入灵活,可批量可随机;
3、管式反应体系,检测结果一致性好;
4、检测通量大,每小时200个测试;
缺点:
1、人机交互的模块分布在仪器的3个角上,不利于操作;
2、开机状态不能加载更换试剂,需停机才能操作;
检测项目:主要包括肝炎项目、肿瘤项目、肝纤 项目、甲状腺项目、心肌标志物项目、炎症项目、 糖代谢项目、高血压项目等。
参考资料:http://wenku.baidu.com/link?url=NtlVCu_jrB2SjBcVKbSZlqC6-c_MiM6tD80PSpMA4fr117cKGqCIujIfWMtKgYS02JWDtDcCLfBMzDIcG0ap1YKKKu2Hdf9qELF-CnZBay3
因为我公司里面臭氧管道是正方形,估计是50cm*50cm哪么大,都不知道怎么取样。求各位高手指点一下。我的QQ是393593653.希望大家帮帮我。
谢谢。
CSY-E96D动物疾病快速诊断仪采用固相酶联免疫吸附ELISA的原理、检验检疫单位使用,广泛应用于养殖场,即酶联免疫法、屠宰场、肉产品深加工企业、猪瘟、猪蓝耳。2着都是利用酶联免疫吸附法ELISA试剂取检测的
是将具有高灵敏度的化学发光测定技术与高特异性的免疫反应相结合,用于各种抗原、半抗原、抗体、激素、酶、脂肪酸、维生素和药物等的检测分析技术。是继放免分析、酶免分析、荧光免疫分析和时间分辨荧光免疫分析之后发展起来的一项最新免疫测定技术。
电化学发光检测器是在高效液相(HPLC)终点上装上电化学反应器,在此反应器中会同时打入一些反应物,使被测物在电化学三电极之下,电化学发生,而被测光的量,再被转换成浓度讯号。
化学发光则不需要电化学的那套系统。在反应器中会同时打入一些反应物,当物质与反应物作用,因化学上的自身氧化还原反应,致使发光。而同时被测到光的量,也是再被转换成浓度讯号。
核酸是一类生物大分子,在蛋白质的合成和遗传中起着重要作用.核酸传感器,特别是DNA传感器的研制引起广泛关注.DNA传感器一股以固定化的单链DNA为分子识别物质.对目标DNA片段(靶序列)进行识别,通过一定的检测手段进行
帮帮忙,谢谢
化学发光免疫分析包括三大类型:即标记化学发光物质的化学发光免疫分析;标记荧光物质的荧光化学发光免疫分析和标记酶的化学发光酶联免疫分析。下面以偶合放大化学发光酶联免疫分析法检测人血清中乙型肝炎表面抗原(HBsAg)为例。
(一) 原理
尽管辣根过氧化物酶(HRP)可以催化Luminol-H2O2反应体系产生化学发光,但由于该体系的检测灵敏度不够高,不能满足酶联免疫测定的要求。因此,为了提高体系的检测灵敏度,可将HRP催化H2O2氧化曙红(Eosin)的反应与该反应产物增强HRP催化luminol-H2O2的化学发光反应相偶合,建立偶合放大化学发光酶联免疫分析法。这里,酶的活性是基于下列发光反应进行检测的:
HRP
luminol+H2O2───→产物+hν
产 物
↑
Eosin+H2O2──────┘
HRP
二) 操作步骤
1. 包被抗体 在每个小试管中加入聚苯乙烯珠各一枚,再加入300μl用0.05M,PH9.6碳酸盐缓冲液稀释的抗HBsAg抗体,同时设空白对照,置4℃过夜。
2. 洗涤 用抽滤针头吸干管内液体,加入Tris-HCl-Tween20洗涤3次,每次加2ml,放置3~5min,用抽滤针头吸干管内液体。
3. 加待检血清和阳性标准品 用PBS-Tween20缓冲液不同倍数稀释HBsAg阳性标准品或待检血清,每管加入300μl。同时设阴性对照;空白对照管只加抗体稀释液。置37℃孵育2h。
4. 洗涤 同2。
5. 加酶标抗体 用含小牛血清的PBS-Tween20缓冲液稀释HRP标记的抗HBsAg抗体,每管加入300μl,空白对照管只加用于稀释酶标抗体的稀释液。置37℃孵育2h。
6. 洗涤 同2。
7. 化学发光测定 给每管加入300μl底物溶液,置37℃保温20min。犎;后将小试放入LKB-1250lumimeter中,并置于测量位置,加入300μl5.0×10-4Mluminol。记录仪记录化学发光强度。
8. 同时用ELISA方法进行对照,结果测量采用DG3022型酶联免疫检测仪。
结果判定
(1) 定性 按下列公式判别阴、阳性:
L样品-L空白 ┌≥2.1 为阳性
S/N=──────── = 商│
L阴性对照-L空白 └<2.1 为阴性
(2) 定量 以不同稀释度的HBsAg阳性标准品的化学发光强度为纵坐标,不同稀释倍数为横坐标,作出剂量反应曲线(标准曲线),犜r待测样品中HBsAg的含量就可由测量的化学发光强度换算得到。
(三) 试剂和器材
1. 试剂
(1) 缓冲液 a. 0.05M,PH9.6Na2CO3-NaHCO3缓冲液(包被液)
b.0.01M,PH7.4PBS-Tween20缓冲液(抗体稀释液)
c.0.02M,PH7.4Tris-HCl-Tween20缓冲液(洗涤液)
(2) 抗体 a.抗HBsAg抗体
b.HRP标记的抗HBsAg抗体
(3) 抗原 a.正常人血清(HBsAg阴性对照)
b.HBsAg阳性标准品
c.待检血清
(4) 小牛血清
(5) 底物溶液
1.0mlEDTA(1.0×10-2M)
2.0mlEosin(1.0×10-3M)
1.0mlH2O2(7.5×10-3M) 用三蒸水稀释到25ml
0.4mlHCl(1.0×10-2M)
0.2mlTween20(1%)
(6) luminol5.0×10-4M
2. 器材
(1) LKB-1250lumimeter
(2)隔水式电热恒温培养箱
(3) 各种规格加样器,小玻璃试管,聚苯乙烯珠
(4) 洗瓶、抽滤装置等
(四) 注意事项
1. 洗涤要彻底,以免因血清中其他来源的过氧化物酶类物质所产生的非特异性反
应,而影响测定结果。
2. 实验中,应分别设置阳性、阴性、空白对照来控制实验条件,且每份样品均应做三个复管,以保证实验结果的准确性。
3. 为了克服酶标抗体因非特异性吸附而造成的较高本底,可用适量小牛血清加以抑制。
4. 当加入luminol后,迅速产生化学发光并使发光在一秒钟内达到峰值,犎;后很快衰减到基线水平。因此,只有当小试管置于仪器的测量位置时,方可加入luminol。
5. 底物的加入,是为了增强化学发光强度。但只有当底物分子与酶催化活性中心充分接触时,反应速度才能加快。当反应进行15min达到平衡时,牷/学发光强度则不再随时间的延长而变化,且在1h内保持稳定,因此,控制底物与酶反应15min后加luminol进行化学发光测定。

