Revision 12

#26369

Store at +4C

CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, 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 Usage Information

Application Dilution
Western Blotting 1:1000
IHC Leica Bond 1:400 - 1:1600
Immunohistochemistry (Paraffin) 1:200 - 1:800
Immunofluorescence (Immunocytochemistry) 1:1600 - 1:6400

Storage

Supplied in PBS. This product is stable for 12 months when stored at -20C. This product will freeze at -20°C so it is recommended to aliquot into single-use vials to avoid multiple freeze/thaw cycles. A slight precipitate may be present, but will not interfere with antibody performance.

Specificity/Sensitivity

SARS-CoV-2 Nucleocapsid Protein (HL344) 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) proteins from SARS or MERS coronaviruses.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a full-length SARS-CoV-2 nucleocapsid (N) recombinant 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 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).

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

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

Cross-Reactivity Key

Vir: Virus

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

SignalStain is a registered trademark of Cell Signaling Technology, Inc.

XP is a registered trademark of Cell Signaling Technology, Inc.

U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

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

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 12

Cell Signaling Technology Logo
Western blot analysis of extracts from 293T cells, mock transfected (lane 1) or transiently transfected with plasmid encoding Myc/DDK-tagged SARS-CoV-2 nucleocapsid (N) protein (lane 2), using SARS-CoV-2 Nucleocapsid Protein (HL344) Rabbit mAb (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western Blotting Image 1: SARS-CoV-2 Nucleocapsid Protein (HL344) Rabbit mAb
Western blot analysis of extracts from transgenic A549 cells stably expressing HA-tagged SARS-CoV-2 nucleocapsid (N) protein (A549-N protein) (lane 1) or HA-tagged SARS-CoV-2 envelope (E) protein (A549-E protein) (lane 2), using SARS-CoV-2 Nucleocapsid Protein (HL344) Rabbit mAb (upper), HA-Tag (C29F4) Rabbit mAb #3724 (middle), and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Absence of signal in A549-E protein cell extracts confirms specificity of the antibody for SARS-CoV-2 nucleocapsid (N) protein. Transgenic A549 cells were generously provided by Dr. Elena Piskounova, Weill Cornell Medicine.
Western Blotting Image 2: SARS-CoV-2 Nucleocapsid Protein (HL344) Rabbit mAb
Immunohistochemical analysis of paraffin-embedded SARS-CoV-2 positive human lung using SARS-CoV-2 Nucleocapsid Protein (HL344) Rabbit mAb performed on the Leica® BOND™ Rx.
Immunohistochemistry Image 1: SARS-CoV-2 Nucleocapsid Protein (HL344) Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 12

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded 293T cell pellet transfected with SARS-CoV-2 nucleocapsid (N) protein (left-top) and various paraffin-embedded human tissues: normal lung (middle-top), normal colon (right-top), normal heart (left-bottom), renal cell carcinoma (middle-bottom), and non-small cell lung carcinoma (right-bottom) using SARS-CoV-2 Nucleocapsid Protein (HL344) Rabbit mAb. Note the lack of staining in the tissues, all of which were procured prior to 2019, and therefore serve as reliable negative controls.
Immunohistochemistry Image 1: SARS-CoV-2 Nucleocapsid Protein (HL344) Rabbit mAb
Immunohistochemical analysis of paraffin-embedded 293T cell pellets, control (left) or SARS-CoV-2 nucleocapsid (N) protein transfected (right), using SARS-CoV-2 Nucleocapsid Protein (HL344) Rabbit mAb.
Immunohistochemistry Image 2: SARS-CoV-2 Nucleocapsid Protein (HL344) Rabbit mAb
Immunohistochemical analysis of paraffin-embedded cell pellets of transgenic A549 cells stably expressing SARS-CoV-2 nucleocapsid (N) protein (A549-N protein) (left, positive) or SARS-CoV-2 envelope (E) protein (A549-E protein) (right, negative), using SARS-CoV-2 Nucleocapsid Protein (HL344) Rabbit mAb. Transgenic A549 cells were generously provided by Dr. Elena Piskounova, Weill Cornell Medicine.
Immunohistochemistry Image 3: SARS-CoV-2 Nucleocapsid Protein (HL344) Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 12

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded SARS-CoV-2-infected hACE2 K18 mouse lung using SARS-CoV-2 Nucleocapsid Protein (HL344) Rabbit mAb. Human ACE2 transgenic mouse lung tissue was generously provided by Dr. Nicholas Crossland and Dr. Florian Douam, National Emerging Infectious Diseases Laboratories, Boston University.
Immunohistochemistry Image 4: SARS-CoV-2 Nucleocapsid Protein (HL344) Rabbit mAb
Immunohistochemical analysis of paraffin-embedded A549-hACE2-hTMPRSS2 cell pellet, mock infected or infected with SARS-CoV-2 2019-nCoV/USA-WA1/2020 variant, SARS-CoV-2 Alpha (B.1.1.7) variant, SARS-CoV-2 Beta (B.1.351) variant (from top left to right as indicated), SARS-CoV-2 Gamma (P.1) variant, SARS-CoV-2 Delta (B.1.617.2) variant, or SARS-CoV-2 Omicron (B.1.1.529) variant (from bottom left to right as indicated), using SARS-CoV-2 Nucleocapsid Protein (HL344) Rabbit mAb. A549-hACE2-hTMPRSS2 cell pellets were generously provided by Dr. Nicholas Crossland and Dr. Florian Douam, National Emerging Infectious Diseases Laboratories, Boston University.
Immunohistochemistry Image 5: SARS-CoV-2 Nucleocapsid Protein (HL344) Rabbit mAb
Confocal immunofluorescent analysis of 293T cells transiently transfected with SARS-CoV-2 nucleocapsid (N) protein, using SARS-CoV-2 Nucleocapsid Protein (HL344) Rabbit mAb (green), DyLight™ 554 Phalloidin #13054 (red), and DAPI #4083 (blue).
Immunofluorescence Image 1: SARS-CoV-2 Nucleocapsid Protein (HL344) Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 12

Cell Signaling Technology Logo
Confocal immunofluorescent analysis of transgenic A549 cells stably expressing SARS-CoV-2 nucleocapsid (N) protein (A549-N protein) (left, positive) and SARS-CoV-2 envelope (E) protein (A549-E protein) (right, negative), using SARS-CoV-2 Nucleocapsid Protein (HL344) Rabbit mAb, DyLight™ 554 Phalloidin #13054 (red), and DAPI #4083 (blue). Transgenic A549 cells were generously provided by Dr. Elena Piskounova, Weill Cornell Medicine.
Immunofluorescence Image 2: SARS-CoV-2 Nucleocapsid Protein (HL344) Rabbit mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.