Revision 5
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.
Applications:

WB, IHC-Bond, IHC-P, IF-IC

REACTIVITY:

Vir

SENSITIVITY:

Endogenous

MW (kDa):

55

Source/Isotype:

Rabbit IgG

UniProt ID:

#P0DTC9

Entrez-Gene Id:

43740575

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
IHC Leica Bond 1:50 - 1:200
Immunohistochemistry (Paraffin) 1:50
Immunofluorescence (Immunocytochemistry) 1:100 - 1:400

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

SARS-CoV-2 Nucleocapsid Protein (E9L7H) XP® Rabbit mAb recognizes endogenous levels of total SARS-CoV-2 nucleocapsid (N) protein. The antibody has been confirmed to detect the following SARS-CoV-2 isolates and variants: USA-WA1/2020, Alpha (B.1.1.7), Beta (B.1.351), Gamma (P.1), Delta (B.1.617.2), and Omicron (B.1.1.529). The antibody does not cross-react with nucleocapsid (N) protein from SARS-CoV-1 coronavirus.

Species Reactivity:

Virus

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the amino terminus of SARS-CoV-2 nucleocapsid (N) protein (USA-WA1/2020 isolate).

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 relatively large and encodes up to 29 open reading frames (ORFs). These include ORF1a and ORF1b (further processed into 16 non-structural proteins), 9 accessory proteins, and 4 canonical structural proteins: spike (S), envelope (E), membrane (M), and nucleocapsid (N) (2). The nucleocapsid (N) protein is a phosphoprotein that plays a critical role in viral assembly (3,4). It binds to the viral genomic RNA, enabling its packaging into the ribonucleoprotein complex (RNP), and is the most abundant protein in the virion (5). Evidence from studies in SARS-CoV-2 and other coronaviruses suggest functions for nucleocapsid (N) protein beyond viral RNA packaging, including roles in viral replication (6) and suppressing host immune responses to viral infection (7).

  1. Zhou, P. et al. (2020) Nature 579, 270-273.
  2. Tortorici, M.A. and Veesler, D. (2019) Adv Virus Res 105, 93-116.
  3. Zengel, J. et al. (2015) J Virol 89, 7338-47.
  4. Surjit, M. et al. (2005) J Virol 79, 11476-86.
  5. Peng, Y. et al. (2020) EMBO J, e105938.
  6. Cong, Y. et al. (2020) J Virol 94, e01925-19. doi: 10.1128/JVI.01925-19.
  7. Mu, J. et al. (2020) Sci China Life Sci 63, 1-4.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IHC-Bond: IHC Leica Bond IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry)

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.
XP is a registered 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
#86326

SARS-CoV-2 Nucleocapsid Protein (E9L7H) XP® Rabbit mAb

Western Blotting Image 1: SARS-CoV-2 Nucleocapsid Protein (E9L7H) XP® Rabbit mAb Expand Image
使用 SARS-CoV-2 Nucleocapsid Protein (E9L7H) XP® Rabbit mAb(上图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对模拟感染(泳道 1)或 SARS-CoV-2 感染(泳道 2)的 Vero-E6 细胞提取物进行蛋白质印迹法分析。SARS-CoV-2 感染的 Vero-E6 细胞由波士顿大学国家新兴传染病实验室 Mohsan Saeed 博士友情提供。
Western Blotting Image 2: SARS-CoV-2 Nucleocapsid Protein (E9L7H) XP® Rabbit mAb Expand Image
使用 SARS-CoV-2 Nucleocapsid Protein (E9L7H) XP® Rabbit mAb(上图)、HA-Tag (6E2) Mouse mAb #2367(中图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对稳定表达 HA 标记的 SARS-CoV-2 核衣壳 (N) 蛋白(A549-N 蛋白)(泳道 1)或 HA 标记的 SARS-CoV-2 包膜 (E) 蛋白(A549-E 蛋白)(泳道 2)的转基因 A549 细胞的提取物进行蛋白质印迹法分析。A549-E 蛋白细胞提取物中信号的缺失证实了该抗体对 SARS-CoV-2 核衣壳(N)蛋白的特异性。转基因 A549 细胞由 Weill Cornell Medicine 的 Elena Piskounova 博士友情提供。
Western Blotting Image 3: SARS-CoV-2 Nucleocapsid Protein (E9L7H) XP® Rabbit mAb Expand Image
使用 SARS-CoV-2 Nucleocapsid Protein (E9L7H) XP® Rabbit mAb(上图)、Myc-Tag (71D10) Rabbit mAb #2278(中图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对转染空载(泳道 1)或瞬时转染带血浆编码的 MycDDK 标记的 SARS-CoV-2 核衣壳 (N) 蛋白(泳道 2)的 293T 细胞提取物进行蛋白质印迹法分析。
Immunohistochemistry Image 1: SARS-CoV-2 Nucleocapsid Protein (E9L7H) XP® Rabbit mAb Expand Image
在 Leica BOND Rx 上使用 SARS-CoV-2 Nucleocapsid Protein (E9L7H) XP® Rabbit mAb 对石蜡包埋的 SARS-CoV-2 阳性人胎盘进行免疫组织化学分析。
Immunohistochemistry Image 1: SARS-CoV-2 Nucleocapsid Protein (E9L7H) XP® Rabbit mAb Expand Image
使用 SARS-CoV-2 Nucleocapsid Protein (E9L7H) XP® Rabbit mAb 对石蜡包埋的 SARS-CoV-2 阳性人肺脏进行免疫组织化学分析。
Immunohistochemistry Image 2: SARS-CoV-2 Nucleocapsid Protein (E9L7H) XP® Rabbit mAb Expand Image
以浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(右图)为对照,使用 SARS-CoV-2 Nucleocapsid Protein (E9L7H) XP® Rabbit mAb(左图)对石蜡包埋的 SARS-CoV-2 感染的 hACE2 K18 小鼠肺脏进行免疫组织化学分析。人ACE2 转基因小鼠肺组织由波士顿大学国家新兴传染病实验室 Nicholas Crossland 博士和 Florian Douam 博士友情提供。
Immunohistochemistry Image 3: SARS-CoV-2 Nucleocapsid Protein (E9L7H) XP® Rabbit mAb Expand Image
使用 SARS-CoV-2 Nucleocapsid Protein (E8R1L) Mouse mAb 对用 SARS-CoV-2 核衣壳蛋白(N)转染且石蜡包埋的以下 293T 细胞沉淀物和各种石蜡包埋的人体组织进行免疫组织化学分析:使用 SARS-CoV-2 Nucleocapsid Protein (E9L7H) XP® Rabbit mAb 对结肠腺癌组织(中上图)、大细胞神经内分泌肺癌组织(右上图)、正常肺脏组织(左下图)、套细胞淋巴瘤组织(中下图)和正常小肠组织(右下图)进行分析。注意组织中没有染色,所有染色均在 2019 之前完成,因此可作为可靠的阴性对照。
Immunohistochemistry Image 4: SARS-CoV-2 Nucleocapsid Protein (E9L7H) XP® Rabbit mAb Expand Image
使用 SARS-CoV-2 Nucleocapsid Protein (E9L7H) XP® Rabbit mAb 对石蜡包埋的对照(左图)或 SARS-CoV-2 核衣壳 (N) 蛋白转染(右图)的 293T 细胞沉淀物进行免疫组织化学分析。
Immunohistochemistry Image 5: SARS-CoV-2 Nucleocapsid Protein (E9L7H) XP® Rabbit mAb Expand Image
使用 SARS-CoV-2 Nucleocapsid Protein (E9L7H) XP® Rabbit mAb 对稳定表达 SARS-CoV-2 核衣壳 (N) 蛋白(A549-N 蛋白)(左图,阳性)或 SARS-CoV-2 包膜 (E) 蛋白(A549-E 蛋白)(右图,阴性)的石蜡包埋的转基因 A549 细胞的细胞沉淀物进行免疫组织化学分析。转基因 A549 细胞由 Weill Cornell Medicine 的 Elena Piskounova 博士友情提供。
Immunohistochemistry Image 6: SARS-CoV-2 Nucleocapsid Protein (E9L7H) XP® Rabbit mAb Expand Image
使用 SARS-CoV-2 Nucleocapsid Protein (E9L7H) XP® Rabbit mAb 对石蜡包埋的模拟感染或 SARS-CoV-2 2019-nCoV/USA-WA1/2020 变体、SARS-CoV-2 Alpha (B.1.1.7) 变体、SARS-CoV-2 Beta (B.1.351) 变体(如图所示,从左上图到右图)、SARS-CoV-2 Gamma (P.1) 变体、SARS-CoV-2 Delta (B.1.617.2) 变体、或 SARS-CoV-2 Omicron (B.1.1.529) 变体感染(如图所示,从左下图到右图)的 A549-hACE2-hTMPRSS2 细胞沉淀物进行免疫组织化学分析。A549-hACE2-hTMPRSS2 细胞沉淀物由波士顿大学国家新兴传染病实验室的 Nicholas Crossland 博士和 Florian Douam 博士慷慨提供。
Immunofluorescence Image 1: SARS-CoV-2 Nucleocapsid Protein (E9L7H) XP® Rabbit mAb Expand Image
使用 SARS-CoV-2 Nucleocapsid Protein (E9L7H) XP® Rabbit mAb(绿色)、DyLight 650 Phalloidin #12956(红色)和 DAPI #4083(蓝色)对稳定表达 SARS-CoV-2 核衣壳 (N) 蛋白(A549-N 蛋白)(左图,阳性)和 SARS-CoV-2 包膜 (E) 蛋白(A549-E 蛋白)(右图,阴性)的转基因 A549 细胞进行共聚焦免疫荧光分析。转基因 A549 细胞由 Weill Cornell Medicine 的 Elena Piskounova 博士友情提供。