Revision 5

#58873Store at +4C

1 个试剂盒

(96 assays)

Species Cross Reactivity

H

UniProt ID:

#P0DTC2

Entrez-Gene Id:

#43740568

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Color Storage Temp
Spike Protein Coated Microwells II 87455 96 tests +4C
Anti-Human IgA, HRP-linked Antibody (ELISA Formulated) 27368 1 ea Red (Lyophilized) +4C
Sample Diluent A 71637 25 ml +4C
HRP Diluent 13515 11 ml Red +4C
ELISA Wash Buffer (20X) 9801 25 ml +4C
TMB Substrate 7004 11 ml +4C
STOP Solution 7002 11 ml +4C
Sealing Tape 54503 2 ea +4C
ELISA Kit #58873 Positive Control 68093 1 ea +4C
ELISA Kit #58873 Negative Control 47262 1 ea +4C

*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.

Description

The SARS-CoV-2 Spike Protein Serological IgA ELISA Kit is a solid-phase ELISA that detects binding of human IgA to full-length ectodomain SARS-CoV-2 spike trimeric protein (S-protein). Trimeric spike protein has been coated onto microwells. After incubation with sample, human IgA specific for trimeric spike protein is captured on the plate. The wells are then washed to remove unbound material. Anti-Human IgA, HRP-linked antibody is then used to recognize the bound IgA. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of IgA specific for spike protein.

*Antibodies in this kit are custom formulations specific to kit.

Specificity/Sensitivity

The SARS-CoV-2 Spike Protein Serological IgA ELISA Kit detects endogenous levels of human IgA directed to full-length ectodomain SARS-CoV-2 spike trimeric protein (S-protein).

Background

The cause of the COVID-19 pandemic is a novel and highly pathogenic coronavirus, termed SARS-CoV-2 (severe acute respiratory syndrome coronavirus-2). SARS-CoV-2 is a member of the Coronaviridae family of viruses (1). The genome of SARS-CoV-2 is similar to other coronaviruses, and is comprised of four key structural proteins: S, the spike protein, E, the envelope protein, M, the membrane protein, and N, the nucleocapsid protein (2). Coronavirus spike proteins are class I fusion proteins and harbor an ectodomain, a transmembrane domain, and an intracellular tail (3,4). The highly glycosylated ectodomain projects from the viral envelope surface and facilitates attachment and fusion with the host cell plasma membrane. The ectodomain can be further subdivided into host receptor-binding domain (RBD) (S1) and membrane-fusion (S2) subunits, which are produced upon proteolysis by host proteases at S1/S2 and S2’ sites. S1 and S2 subunits remain associated after cleavage and assemble into crown-like homotrimers (2,4). In humans, both SARS-CoV and SARS-CoV-2 spike proteins utilize the angiotensin-converting enzyme 2 (ACE2) protein as a receptor for cellular entry (5-7). Spike protein subunits represent a key antigenic feature of coronavirus virions, and therefore represent an important target of vaccines, novel therapeutic antibodies, and small-molecule inhibitors (8,9).

  1. Zhou, P. et al. (2020) Nature 579, 270-3.
  2. Tortorici, M.A. and Veesler, D. (2019) Adv Virus Res 105, 93-116.
  3. Li, F. et al. (2006) J Virol 80, 6794-800.
  4. Li, F. (2016) Annu Rev Virol 3, 237-61.
  5. Shang, J. et al. (2020) Nature 581, 221-4.
  6. Wrapp, D. et al. (2020) Science 367, 1260-3.
  7. Yan, R. et al. (2020) Science 367, 1444-8.
  8. Yuan, Y. et al. (2017) Nat Commun 8, 15092.
  9. Amanat, F. and Krammer, F. (2020) Immunity 52, 583-9.

Background References

    Cross-Reactivity Key

    H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    限制使用

    除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

    专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

    Revision 5
    #58873

    SARS-CoV-2 Spike Protein Serological IgA ELISA Kit

    SARS-CoV-2 Spike Protein Serological IgA ELISA Kit: Image 1 Expand Image
    患者测试:使用 SARS-CoV-2 Spike Protein Serological IgA ELISA Kit #58873 检测患者样品。从诊断为 SARS-CoV-2 的供体(诊断阳性,n=59*),或从 SARS-CoV-2 爆发之前收集的假定未感染供体(假定阴性,n=86 )中获得血清或血浆。如实验步骤中所述,在运行检测之前将样品加热灭活(56°C,30 分钟)并以 1:200 的比例稀释。根据试剂盒实验步骤的“数据分析”部分中所述的标准,样品可视为阳性、阴性或不确定。从这些数据计算出阳性符合率 (PPA) 和阴性符合率 (NPA)。

    *阳性参比样本来自患有阳性 SARS-CoV-2 诊断的患者供体。但是,SARS-CoV-2 阳性诊断并不总是与 IgA 血清学 ELISA 阳性反应相关,因为相对于疾病发作,疾病严重程度和样品采集时间的差异会影响参考样品中 IgG、IgA 和 IgM 的存在和丰度。

    注:我们将继续检测更多样品(如可用)。如需获取最新数据集,请始终参考网站产品页面上的 #58873。

    SARS-CoV-2 Spike Protein Serological IgA ELISA Kit: Image 2 Expand Image
    测定内精度:使用一个 SARS-CoV-2 Spike Protein Serological IgA ELISA #58873 测定试剂盒,分别对三个不同的血清样品进行 24 次重复测试。计算每个样品的测定内 CV (%),并且使用所附实验步骤中所述的临界标准与阴性对照进行比较时,每个复孔样品都被正确地标识为阳性或阴性。
    SARS-CoV-2 Spike Protein Serological IgA ELISA Kit: Image 3 Expand Image
    测定内精度:使用 3 个不同血清样品设置复孔,阳性和阴性对照设立 4 个复孔,在同一批次的六个测定试剂盒中检测。计算每个样品的分析间 CV (%),并且使用所附实验步骤中所述的临界标准与阴性对照进行比较时,每个测定试剂盒均正确地将样品鉴定为阳性或阴性。
    SARS-CoV-2 Spike Protein Serological IgA ELISA Kit: Image 4 Expand Image
    患者测试:使用 SARS-CoV-2 Spike Protein Serological IgA ELISA Kit #58873 检测患者样品。如实验步骤中所述,在运行检测之前将血清/血浆样品加热灭活(56°C,30 分钟)并以 1:200 的比例稀释。绘制了各血清/血浆样品在 450 nm 处的背景减去吸光度的值,与获自诊断为 SARS-COV-2(诊断阳性,n=59*)的供体的样品相对应的值绘制于右侧,与来自 SARS-CoV-2 爆发之前收集的假定未感染供体(假定阴性,n= 86)的样品相对应的值绘制于左侧。同时也显示了分界线(蓝线),位于分界线(蓝线)之上的样品视为阳性,以及阴性分界线(红线),位于阴性分界线(红线)之下的样品视为阴性。截止值按照试剂盒实验步骤的“数据分析”部分所述进行计算。

    *阳性参比样本来自患有阳性 SARS-CoV-2 诊断的患者供体。但是,SARS-CoV-2 阳性诊断并不总是与 IgA 血清学 ELISA 阳性反应相关,因为相对于疾病发作,疾病严重程度和样品采集时间的差异会影响参考样品中 IgG、IgA 和 IgM 的存在和丰度。