Revision 1

#34340Store at -20C

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, IP

REACTIVITY:

Vir

SENSITIVITY:

Endogenous

MW (kDa):

25-31

SOURCE:

Rabbit

UniProt ID:

#P0DTC3

Entrez-Gene Id:

43740569

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50

Storage

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

Specificity / Sensitivity

SARS-CoV-2 ORF3a Antibody recognizes endogenous levels of total SARS-CoV-2 ORF3a protein.

Species Reactivity:

Virus

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ile232 of SARS-CoV-2 ORF3a protein. Antibodies are purified by protein A and peptide affinity chromatography.

Background

SARS-CoV-2 ORF3a is the largest of the SARS-CoV-2 accessory proteins containing three amino-terminal transmembrane domains localized to the plasma membrane and perinuclear regions including the golgi apparatus (1). It contains several functional domains including a TRAF3 binding motif, a caveolin-binding motif, a tyrosine-sorting motif, and a diacidic motif (2). SARS-CoV ORF3a was shown to regulate apoptotic and necrotic cell death and inhibit late stages of autophagy by inhibiting fusion of the autophagosome and lysosome (3-5). SARS-CoV-2 ORF3a was also found to inhibit late stages of autophagy by inhibiting fusion of the autophagosome and lysosome through association with components of the homotypic fusion and protein sorting HOPS complex (6-8). It is thought that SARS-CoV-2 can form ion channels that regulates viral release (9). In addition, it can regulate several aspects of innate immune responses, including TRAF3 signaling, activation of NF-κB, and production of inflammatory cytokines (10,11).
α

  1. Redondo, N. et al. (2021) Front Immunol 12, 708264.
  2. Issa, E. et al. (2020) mSystems 5, e00266-20. doi: 10.1128/mSystems.00266-20.
  3. Yue, Y. et al. (2018) Cell Death Dis 9, 904.
  4. Ren, Y. et al. (2020) Cell Mol Immunol 17, 881-883.
  5. Law, P.T.W. et al. (2005) J Gen Virol 86, 1921-1930.
  6. Zhang, Y. et al. (2021) Cell Discov 7, 31.
  7. Miao, G. et al. (2021) Dev Cell 56, 427-442.e5.
  8. Qu, Y. et al. (2021) Front Cell Dev Biol 9, 716208.
  9. Marquez-Miranda, V. et al. (2020) bioRxiv, 10.22.349522. doi: 10.1101/2020.10.22.349522.
  10. Su, C.M. et al. (2021) Sci Rep 11, 13464.
  11. Siu, K.L. et al. (2019) FASEB J 33, 8865-8877.

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 IP: Immunoprecipitation

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 1
#34340

SARS-CoV-2 ORF3a Antibody

Western Blotting Image 1: SARS-CoV-2 ORF3a Antibody Expand Image
使用 SARS-CoV-2 ORF3a Antibody(上图)和 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对模拟感染 (-) 或感染 SARS-CoV-2 (+) 的 Vero-E6 细胞提取物进行蛋白质印迹分析。
Western Blotting Image 2: SARS-CoV-2 ORF3a Antibody Expand Image
使用 SARS-CoV-2 ORF3a Antibody(上图)和 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对转染空载 (-) 或转染带有 Myc/DDK 标记的全长 SARS-CoV-2 ORF3a 蛋白 (SARS-CoV-2 ORF3a-Myc/DDK; +) 表达载体的 293T 细胞提取物进行蛋白质印迹分析。
Immunoprecipitation Image 1: SARS-CoV-2 ORF3a Antibody Expand Image
对转染 Myc/DDK 标记的全长 SARS-CoV-2 ORF3a 蛋白表达载体的 293T 细胞提取物中的 SARS-CoV-2 ORF3a 蛋白进行免疫沉淀分析。泳道 1 是 10% input,泳道 2 是 Normal Rabbit IgG #2729,和泳道 3 为 SARS-CoV-2 ORF3a Antibody。使用 SARS-CoV-2 ORF3a Antibody 进行蛋白质印迹分析。Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 用作二抗。