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AB284634

Anti-SARS-CoV-2 nsp6 antibody [EPR24845-90] - BSA and Azide free

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

View Alternative Names

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

2 Images
Flow Cytometry (Intracellular) - Anti-SARS-CoV-2 nsp6 antibody [EPR24845-90] - BSA and Azide free (AB284634)
  • Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-SARS-CoV-2 nsp6 antibody [EPR24845-90] - BSA and Azide free (AB284634)

Flow cytometric analysis 4% Paraformaldehyde fixed 90% methanol permeabilised HEK-293T (Human embryonic kidney epithelial) cells transfected with a SARS-CoV-2 nsp9 (Left) or SARS-CoV2 nsp6 (Right) expression vector containing a myc tag labelling SARS-CoV-2 nsp6 with ab284633 at 1/500 dilution. Goat anti rabbit IgG (Alexa Fluor®488) (ab150081) at 1/5000 dilution was used as the secondary antibody. This data was developed using ab284633, the same antibody clone in a different buffer formulation.

Immunocytochemistry/ Immunofluorescence - Anti-SARS-CoV-2 nsp6 antibody [EPR24845-90] - BSA and Azide free (AB284634)
  • ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SARS-CoV-2 nsp6 antibody [EPR24845-90] - BSA and Azide free (AB284634)

Immunofluorescent analysis of 4% Paraformaldehyde fixed 0.1% TritonX-100 permeabilised HEK-293T (Human embryonic kidney 293) cells labelling Anti-SARS-CoV-2 nsp6 with ab284633 at 1/500 dilution, followed by AlexaFluor®488 Goat anti-Rabbit secondary antibody (ab150081) at 1/1000 dilution. Confocal image showing positive staining in HEK-293T cells transfected with a SARS-CoV-2 nsp6 expression vector containing a myc tag, and no staining in HEK-293T cells transfected with a SARS-CoV-2 nsp9 expression vector containing a myc tag is observed. Myc-Tag Mouse mab (Alexa Fluor® 647) was used to counterstain Myc-Tag at 1/100 dilution (Red).The Nuclear counterstain is DAPI. This data was developed using ab284633, the same antibody clone in a different buffer formulation.

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR24845-90

Isotype

IgG

Carrier free

Yes

Reacts with

SARS-CoV-2

Applications

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

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"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"}, "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": { "SARS-CoV-2": { "ICCIF-species-checked": "guaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "guaranteed", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-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>Serial dilution range 0.016 - 1000 ng/mL (a 3-fold serial dilution).</p>" }, "Transfected cell line - SARS-CoV-2": { "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>", "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-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

ab284634 is the PBS only version of ab284633.

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.

Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Constituents: 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 nsp6 also known as non-structural protein 6 plays an important role in the lifecycle of the virus. This protein is part of the replicase polyprotein and has an estimated molecular weight of about 33 kDa. It is expressed mainly within infected host cells where it localizes to the endoplasmic reticulum. nsp6 helps in modifying host cell membranes to form vesicles which are necessary for replication and transcription of the viral RNA.
Biological function summary

Non-structural protein 6 is involved in the host cell's autophagy process. It interacts with other non-structural proteins like nsp3 and nsp4 to form a complex that transforms host cell membranes. These membrane structures can shield viral components from the host’s immune system. Autophagy inhibition by nsp6 can also help in providing the virus with a platform for its replication processes.

Pathways

SARS-CoV-2 nsp6 participates in the early vesicle formation process critical to the viral replication complex. It works within the autophagy and endoplasmic reticulum stress pathways. In the autophagy pathway nsp6 connects with proteins such as LC3 and ATG5 which control autophagosome formation. This connection allows the virus to evade cellular mechanisms meant to degrade it enhancing viral persistence and replication in host cells.

SARS-CoV-2 nsp6 relates closely to COVID-19. This protein’s interaction with the autophagy process may contribute to the inflammatory response observed during infection. Moreover its functions link to proteins like nsp3 increasing viral replication which can worsen disease progression. This might lead to severe respiratory conditions and has implications in the development of therapies targeting those protein interactions.

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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