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AB284052

Anti-SARS-CoV-2 nsp16 antibody [EPR24841-18] - BSA and Azide free

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Rabbit Recombinant Monoclonal R1AB antibody. Carrier free. Suitable for Flow Cyt (Intra), ICC/IF, WB, I-ELISA and reacts with Transfected cell line - SARS-CoV-2, Recombinant full length protein - SARS-CoV-2 samples.

View Alternative Names

1a-1b, rep, Replicase polyprotein 1ab, pp1ab, ORF1ab polyprotein

4 Images
Immunocytochemistry/ Immunofluorescence - Anti-SARS-CoV-2 nsp16 antibody [EPR24841-18] - BSA and Azide free (AB284052)
  • ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SARS-CoV-2 nsp16 antibody [EPR24841-18] - BSA and Azide free (AB284052)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) cells labelling SARS-CoV-2 nsp16 with primary antibody anti-SARS-CoV-2 nsp16 (ab284038) at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150081) secondary antibody at 1/1000 dilution. Confocal image showing positive staining in HEK-293T cells transfected with a SARS-CoV2 nsp16 expression vector containing a myc tag, no staining in HEK-293T cells transfected with a SARS-CoV2 nsp9 expression vector containing a myc tag. Myc-Tag Mouse mab (Alexa Fluor® 647) (red) was used to counterstain cells at 1/100 dilution. The nuclear counter stain is DAPI (blue).

This data was developed using ab284038, the same antibody clone in a different buffer formulation.

Flow Cytometry (Intracellular) - Anti-SARS-CoV-2 nsp16 antibody [EPR24841-18] - BSA and Azide free (AB284052)
  • Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-SARS-CoV-2 nsp16 antibody [EPR24841-18] - BSA and Azide free (AB284052)

Intracellular Flow Cytometry analysis of HEK-293T (Human embryonic kidney epithelial cell) cells transfected with a SARS-CoV-2 nsp9 (Left) or SARS-CoV2 nsp16 (Right) expression vector containing a myc tag labeling SARS-CoV-2 nsp16 with ab284038 at 1/500 dilution (Left and Right). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 647, ab150083) secondary antibody was used at 1/5000 dilution.

This data was developed using ab284038, the same antibody clone in a different buffer formulation.

Indirect ELISA - Anti-SARS-CoV-2 nsp16 antibody [EPR24841-18] - BSA and Azide free (AB284052)
  • I-ELISA

Supplier Data

Indirect ELISA - Anti-SARS-CoV-2 nsp16 antibody [EPR24841-18] - BSA and Azide free (AB284052)

This data was developed using ab284038, the same antibody clone in a different buffer formulation.

Indirect ELISA showing primary antibody ab284038 binding to recombinant SARS-CoV-2 NSP10/16 His-tag protein. Plates were coated with recombinant SARS-CoV-2 NSP10/16 His-tag protein and recombinant SARS-COV-2 NSP9 His-tag protein at 1000 ng/ml. Binding of ab284038 was assessed in a serial dilution range 0.016- 1000 ng/mL (a 3-fold serial dilution).

Binding was detected using pre-adsorbed secondary antibody, goat anti-rabbit IgG H&L (HRP, ab97080) at 1/2000 dilution.

Western blot - Anti-SARS-CoV-2 nsp16 antibody [EPR24841-18] - BSA and Azide free (AB284052)
  • WB

Supplier Data

Western blot - Anti-SARS-CoV-2 nsp16 antibody [EPR24841-18] - BSA and Azide free (AB284052)

Lanes 1 - 2 : Red – loading control ab18184 (Mouse monoclonal [HIS.H8] to 6X His tag®) observed at 36 kDa.

Lanes 3 - 4 : Green – ab284038 observed at 36 kDa (NSP16).

ab284038 was shown to bind specifically to SARS-CoV-2 nsp16 protein in Western blot. Samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

Blocking buffer : 3% milk in TBS-0.1% Tween® 20 (TBS-T)

All lanes:

Western blot - Anti-SARS-CoV-2 nsp16 antibody [EPR24841-18] (<a href='/en-us/products/primary-antibodies/sars-cov-2-nsp16-antibody-epr24841-18-ab284038'>ab284038</a>) at 1/1000 dilution

Lanes 1 and 3:

Recombinant SARS-COV-2 NSP10/NSP16 methyltransferase protein (his-tagged) at 0.5 µg

Lanes 2 and 4:

Recombinant SARS-COV-2 NSP10/NSP16 methyltransferase protein (his-tagged) at 0.2 µg

Observed band size: 36 kDa

false

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR24841-18

Isotype

IgG

Carrier free

Yes

Reacts with

SARS-CoV-2

Applications

Flow Cyt (Intra), ICC/IF, I-ELISA, WB

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IELISA" : {"fullname" : "Indirect ELISA", "shortname":"I-ELISA"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Recombinant full length protein - SARS-CoV-2": { "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>", "IELISA-species-checked": "testedAndGuaranteed", "IELISA-species-dilution-info": "", "IELISA-species-notes": "<p></p>" }, "SARS-CoV-2": { "FlowCytIntra-species-checked": "predicted", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "ICCIF-species-checked": "predicted", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "WB-species-checked": "predicted", "WB-species-dilution-info": "", "WB-species-notes": "", "IELISA-species-checked": "predicted", "IELISA-species-dilution-info": "", "IELISA-species-notes": "" }, "Transfected cell line - SARS-CoV-2": { "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "<p></p>", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "IELISA-species-checked": "notRecommended", "IELISA-species-dilution-info": "", "IELISA-species-notes": "" } } }

Product details

ab284052 is the carrier-free version of ab284038.

Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.4 Constituents: 100% PBS
Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

SARS-CoV-2 nsp16 also known as non-structural protein 16 is an important enzyme in the replication and transcription machinery of the virus. It exhibits 2'-O-methyltransferase activity and has a molecular mass of approximately 33 kDa. Nsp16 is expressed in host cells once the virus infects the cell and translates its polyproteins. Its main role is to modify the cap structure of viral mRNA which helps in avoiding host immune detection and promoting efficient viral replication.
Biological function summary

The function of nsp16 centers around its interaction with nsp10 forming a stable 2'-O-methyltransferase complex. This enzyme complex is responsible for the methylation of the ribonucleic acid cap at the first nucleotide. This cap modification is necessary for the stability of the viral mRNA and its ability to be recognized by the host's cellular machinery as native mRNA which prevents antiviral defenses from targeting the virus. The methylation activity of nsp16 is highly specific requiring both nsp10 and nsp16 to function together effectively.

Pathways

Nsp16 plays an integral role in the viral replication-transcription complex (RTC) pathway. This pathway is essential for the synthesis of viral genomic and subgenomic RNAs facilitating the life cycle of SARS-CoV-2. The RTC includes other viral proteins like nsp12 which acts as the RNA-dependent RNA polymerase. These interactions ensure the replication fidelity and quantity necessary for viral protein production and spreading within the host organism.

Nsp16 is connected to COVID-19 the disease caused by SARS-CoV-2 infection. Its role in immune evasion makes it a target for antiviral drug development as disabling the nsp16 function could make the virus more susceptible to host immune responses. nsp16's cooperation with nsp10 is also significant in this context as disrupting their interaction might prevent the virus from effectively replicating offering a potential therapeutic strategy to combat COVID-19.

Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:

Target data

Replicase polyprotein 1ab. Multifunctional protein involved in the transcription and replication of viral RNAs. Contains the proteinases responsible for the cleavages of the polyprotein.. Host translation inhibitor nsp1. Inhibits host translation by associating with the open head conformation of the 40S subunit (PubMed : 32680882, PubMed : 32908316, PubMed : 33080218, PubMed : 33479166). The C-terminus binds to and obstructs ribosomal mRNA entry tunnel (PubMed : 32680882, PubMed : 32908316, PubMed : 33080218, PubMed : 33479166). Thereby inhibits antiviral response triggered by innate immunity or interferons (PubMed : 32680882, PubMed : 32979938, PubMed : 33080218). The nsp1-40S ribosome complex further induces an endonucleolytic cleavage near the 5'UTR of host mRNAs, targeting them for degradation (By similarity). This inhibits the integrated stress response (ISR) in the infected cell by preventing EIF2S1/eIF2-alpha phosphorylation upstream of stress granule formation and depletes host G3BP1 (PubMed : 36534661). Viral mRNAs less susceptible to nsp1-mediated inhibition of translation, because of their 5'-end leader sequence (PubMed : 32908316, PubMed : 33080218).. Non-structural protein 2. Enhances mRNA repression of the 4EHP-GYF2 complex in the host, thereby inhibiting the antiviral response and facilitating SARS-CoV-2 replication. Possibly acts in cooperation with nsp1, which induces ribosome stalling on host mRNA, triggering mRNA repression by the host 4EHP-GYF2 complex which is enhanced by nsp2.. Papain-like protease nsp3. Responsible for the cleavages located at the N-terminus of the replicase polyprotein. Participates together with nsp4 in the assembly of virally-induced cytoplasmic double-membrane vesicles necessary for viral replication (PubMed : 35551511). Antagonizes innate immune induction of type I interferon by blocking the phosphorylation, dimerization and subsequent nuclear translocation of host IRF3 (PubMed : 32733001). Prevents also host NF-kappa-B signaling (By similarity). In addition, PL-PRO possesses a deubiquitinating/deISGylating activity and processes both 'Lys-48'- and 'Lys-63'-linked polyubiquitin chains from cellular substrates (PubMed : 32726803). Cleaves preferentially ISG15 from antiviral protein IFIH1 (MDA5), but not RIGI (PubMed : 33727702). Can play a role in host ADP-ribosylation by ADP-ribose (PubMed : 32578982). Plays a role in the formation and maintenance of double membrane vesicles (DMVs) replication organelles (PubMed : 35551511). DMVs are formed by nsp3 and nsp4, while nsp6 zippers ER membranes and connects to lipid droplets (PubMed : 35551511).. Non-structural protein 4. Plays a role in the formation and maintenance of double membrane vesicles (DMVs) replication organelles (PubMed : 35551511). DMVs are formed by nsp3 and nsp4, while nsp6 zippers ER membranes and connects to lipid droplets (PubMed : 35551511).. 3C-like proteinase nsp5. Cleaves the C-terminus of replicase polyprotein at 11 sites (PubMed : 32321856). Recognizes substrates containing the core sequence [ILMVF]-Q-|-[SGACN] (PubMed : 32198291, PubMed : 32272481). May cleave human NLRP1 in lung epithelial cells, thereby activating the NLRP1 inflammasome pathway (PubMed : 35594856). May cleave human GSDMD, triggering alternative GSDME-mediated epithelial cell death upon activation of the NLRP1 inflammasome, which may enhance the release interleukins 1B, 6, 16 and 18 (PubMed : 35594856). Also able to bind an ADP-ribose-1''-phosphate (ADRP) (PubMed : 32198291, PubMed : 32272481).. Non-structural protein 6. Plays a role in the formation and maintenance of double membrane vesicles (DMVs) replication organelles (PubMed : 35551511). DMVs are formed by nsp3 and nsp4, while nsp6 zippers ER membranes and connects to lipid droplets (PubMed : 35551511). LDs are consumed during DMV formation (PubMed : 35551511). Binds to host TBK1 without affecting TBK1 phosphorylation; the interaction with TBK1 decreases IRF3 phosphorylation, which leads to reduced IFN-beta production (PubMed : 32979938).. Non-structural protein 7. Plays a role in viral RNA synthesis (PubMed : 32277040, PubMed : 32358203, PubMed : 32438371, PubMed : 32526208). Forms a hexadecamer with nsp8 (8 subunits of each) that may participate in viral replication by acting as a primase. Alternatively, may synthesize substantially longer products than oligonucleotide primers (By similarity).. Non-structural protein 8. Plays a role in viral RNA synthesis (PubMed : 32277040, PubMed : 32358203, PubMed : 32438371, PubMed : 32526208). Forms a hexadecamer with nsp7 (8 subunits of each) that may participate in viral replication by acting as a primase. Alternatively, may synthesize substantially longer products than oligonucleotide primers (By similarity). Interacts with ribosome signal recognition particle RNA (SRP) (PubMed : 33080218). Together with NSP9, suppress protein integration into the cell membrane, thereby disrupting host immune defenses (PubMed : 33080218).. Viral protein genome-linked nsp9. Forms a primer, NSP9-pU, which is utilized by the polymerase for the initiation of RNA chains (PubMed : 37794589). Interacts with ribosome signal recognition particle RNA (SRP) (PubMed : 33080218). Together with NSP8, suppress protein integration into the cell membrane, thereby disrupting host immune defenses (PubMed : 33080218).. Non-structural protein 10. Plays a pivotal role in viral transcription by stimulating both nsp14 3'-5' exoribonuclease (By similarity) and nsp16 2'-O-methyltransferase activities (PubMed : 35944563). Therefore plays an essential role in viral mRNAs cap methylation.. RNA-directed RNA polymerase nsp12. RNA-directed RNA polymerase that catalyzes the transcription of viral genomic and subgenomic RNAs. Acts in complex with nsp7 and nsp8 to transcribe both the minus and positive strands of genomic RNA (PubMed : 32277040, PubMed : 32358203, PubMed : 32438371, PubMed : 32526208). The kinase-like NiRAN domain of NSP12 attaches one or more nucleotides to the amino terminus of NSP9, forming a covalent RNA-protein intermediate that serves as transcription/replication primer (PubMed : 37794589). Subgenomic RNAs (sgRNAs) are formed by discontinuous transcription : The polymerase has the ability to pause at transcription-regulating sequences (TRS) and jump to the leader TRS, resulting in a major deletion (PubMed : 35706445). This creates a series of subgenomic RNAs that are replicated, transcribed and translated (PubMed : 35706445). In addition, Nsp12 is a subunit of the viral RNA capping enzyme that catalyzes the RNA guanylyltransferase reaction for genomic and sub-genomic RNAs (PubMed : 35944563). Subsequently, the NiRAN domain transfers RNA to GDP, and forms the core cap structure GpppA-RNA (PubMed : 35944563).. Helicase nsp13. Plays a role in viral RNA synthesis (PubMed : 33232691). Multi-functional protein with a zinc-binding domain in N-terminus displaying RNA and DNA duplex-unwinding activities with 5' to 3' polarity. Activity of helicase is dependent on magnesium (By similarity). Binds to host TBK1 and inhibits TBK1 phosphorylation; the interaction with TBK1 decreases IRF3 phosphorylation, which leads to reduced IFN-beta production (PubMed : 32979938).. Guanine-N7 methyltransferase nsp14. Plays a role in viral RNA synthesis through two distinct activities. The N7-guanine methyltransferase activity plays a role in the formation of the cap structure GpppA-RNA (PubMed : 35944563). The proofreading exoribonuclease reduces the sensitivity of the virus to RNA mutagens during replication (By similarity). This activity acts on both ssRNA and dsRNA in a 3'-5' direction (By similarity).. Uridylate-specific endoribonuclease nsp15. Plays a role in viral transcription/replication and prevents the simultaneous activation of host cell dsRNA sensors, such as MDA5/IFIH1, OAS, and PKR (By similarity). Acts by degrading the 5'-polyuridines generated during replication of the poly(A) region of viral genomic and subgenomic RNAs (PubMed : 33504779, PubMed : 33564093). Catalyzes a two-step reaction in which a 2'3'-cyclic phosphate (2'3'-cP) is first generated by 2'-O transesterification, which is then hydrolyzed to a 3'-phosphate (3'-P) (PubMed : 33504779, PubMed : 33564093). If not degraded, poly(U) RNA would hybridize with poly(A) RNA tails and activate host dsRNA sensors (By similarity). May bind genomic dsRNA in association with the replication-transcription complex (RTC), and play a role in nsp12 discontinous transcription (PubMed : 34562452, PubMed : 35706445).. 2'-O-methyltransferase nsp16. Methyltransferase that mediates mRNA cap 2'-O-ribose methylation to the 5'-cap structure of viral mRNAs (PubMed : 35944563). N7-methyl guanosine cap is a prerequisite for binding of nsp16 (PubMed : 35944563). Therefore plays an essential role in viral mRNAs cap methylation which is essential to evade immune system (PubMed : 35944563). May disrupt host mRNA splicing in nucleus by interacting with pre-mRNA Recognition Domains ofthe U1 and U2 snRNAs (PubMed : 33080218).
See full target information rep

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