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Rabbit Recombinant Monoclonal R1AB antibody. Suitable for WB, IHC-P, I-ELISA, ICC/IF, Flow Cyt and reacts with SARS-CoV-2, Recombinant full length protein - SARS-CoV-2 samples.


Images

Western blot - Anti-SARS-CoV-2 nsp15 antibody [EPR24840-74] (AB284629), expandable thumbnail
  • Indirect ELISA - Anti-SARS-CoV-2 nsp15 antibody [EPR24840-74] (AB284629), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SARS-CoV-2 nsp15 antibody [EPR24840-74] (AB284629), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SARS-CoV-2 nsp15 antibody [EPR24840-74] (AB284629), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SARS-CoV-2 nsp15 antibody [EPR24840-74] (AB284629), expandable thumbnail

Key facts

Host species

Rabbit

Storage buffer

Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

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

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBIHC-PI-ELISAICC/IFFlow Cyt
Recombinant full length protein - SARS-CoV-2
Not recommended
Tested
Not recommended
Tested
Tested
SARS-CoV-2
Tested
Expected
Tested
Expected
Expected

Tested
Tested

Species

SARS-CoV-2

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Recombinant full length protein - SARS-CoV-2

Dilution info

-

Notes

-

Tested
Tested

Species

Recombinant full length protein - SARS-CoV-2

Dilution info

1/100

Notes

-

Expected
Expected

Species

SARS-CoV-2

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

SARS-CoV-2

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Recombinant full length protein - SARS-CoV-2

Dilution info

-

Notes

-

Tested
Tested

Species

Recombinant full length protein - SARS-CoV-2

Dilution info

-

Notes

-

Expected
Expected

Species

SARS-CoV-2

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Recombinant full length protein - SARS-CoV-2

Dilution info

-

Notes

-

Expected
Expected

Species

SARS-CoV-2

Dilution info

Use at an assay dependent concentration.

Notes

-

Target data

Function

Replicase polyprotein 1abMultifunctional protein involved in the transcription and replication of viral RNAs. Contains the proteinases responsible for the cleavages of the polyprotein.Host translation inhibitor nsp1Inhibits host translation by interacting with binds to the host 40S subunit in ribosomal complexes, including the 43S pre-initiation complex and the non-translating 80S ribosome (PubMed:32680882,PubMed:32908316). The C-terminus binds to and obstructs ribosomal mRNA entry tunnel (PubMed:32680882,PubMed:32908316). Thereby inhibits antiviral response triggered by innate immunity or interferons (PubMed:32680882,PubMed:32979938). The nsp1-40S ribosome complex further induces an endonucleolytic cleavage near the 5'UTR of host mRNAs, targeting them for degradation (By similarity). Viral mRNAs less susceptible to nsp1-mediated inhibition of translation, because of their 5'-end leader sequence (PubMed:32908316). By suppressing host gene expression, nsp1 facilitates efficient viral gene expression in infected cells and evasion from host immune response (By similarity).Non-structural protein 2May play a role in the modulation of host cell survival signaling pathway by interacting with host PHB and PHB2. Indeed, these two proteins play a role in maintaining the functional integrity of the mitochondria and protecting cells from various stresses.Non-structural protein 3Responsible 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 (By similarity). 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 substrates in vitro (PubMed:32726803). Can play a role in host ADP-ribosylation by binding ADP-ribose (PubMed:32578982).Non-structural protein 4Participates in the assembly of virally-induced cytoplasmic double-membrane vesicles necessary for viral replication.3C-like proteinaseCleaves the C-terminus of replicase polyprotein at 11 sites (PubMed:32321856). Recognizes substrates containing the core sequence [ILMVF]-Q-|-[SGACN] (PubMed:32198291, PubMed:32272481). Also able to bind an ADP-ribose-1''-phosphate (ADRP) (By similarity) (PubMed:32198291, PubMed:32272481).Non-structural protein 6Plays a role in the initial induction of autophagosomes from host reticulum endoplasmic (By similarity). Later, limits the expansion of these phagosomes that are no longer able to deliver viral components to lysosomes (By similarity). 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 7Plays a role in viral RNA synthesis (PubMed:32358203, PubMed:32277040, 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 8Plays a role in viral RNA synthesis (PubMed:32358203, PubMed:32277040, 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).Non-structural protein 9May participate in viral replication by acting as a ssRNA-binding protein.Non-structural protein 10Plays a pivotal role in viral transcription by stimulating both nsp14 3'-5' exoribonuclease and nsp16 2'-O-methyltransferase activities. Therefore plays an essential role in viral mRNAs cap methylation.RNA-directed RNA polymeraseResponsible for replication and transcription of the viral RNA genome.HelicaseMulti-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).Proofreading exoribonucleaseEnzyme possessing two different activities: an exoribonuclease activity acting on both ssRNA and dsRNA in a 3' to 5' direction and a N7-guanine methyltransferase activity. Acts as a proofreading exoribonuclease for RNA replication, thereby lowering The sensitivity of the virus to RNA mutagens.Uridylate-specific endoribonucleaseMn(2+)-dependent, uridylate-specific enzyme, which leaves 2'-3'-cyclic phosphates 5' to the cleaved bond.2'-O-methyltransferaseMethyltransferase that mediates mRNA cap 2'-O-ribose methylation to the 5'-cap structure of viral mRNAs. N7-methyl guanosine cap is a prerequisite for binding of nsp16. Therefore plays an essential role in viral mRNAs cap methylation which is essential to evade immune system.

Targets

rep

Alternative names

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

Recommended products

Rabbit Recombinant Monoclonal R1AB antibody. Suitable for WB, IHC-P, I-ELISA, ICC/IF, Flow Cyt and reacts with SARS-CoV-2, Recombinant full length protein - SARS-CoV-2 samples.

Alternative names

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

Key facts

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EPR24840-74

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

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

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.

Supplementary info

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

Activity summary

SARS-CoV-2 nsp15 also known as non-structural protein 15 is an endoribonuclease with uridine-specific activity. It is expressed in the cytoplasm of cells infected by the virus. Nsp15 has a monomer mass of approximately 42 kDa. In its active form it typically assembles into a hexameric complex which is important for its enzymatic function. It helps in evading the host immune response by degrading double-stranded RNA intermediates preventing detection by the host's antiviral pathways.

Biological function summary

Nsp15 is an important participant in the viral replication machinery of SARS-CoV-2. Nsp15 as part of the replication and transcription complex processes viral RNA to promote efficient replication. This protein may interact indirectly with other non-structural proteins such as nsp8 and nsp12 which form part of the RNA-dependent RNA polymerase complex. By maintaining the integrity of viral RNA nsp15 ensures successful virus propagation.

Pathways

Nsp15 plays a significant role in the viral life cycle particularly in the genome replication and transcription pathways. This protein is connected to nsp10 and nsp14 which are involved in RNA proofreading and capping. These interactions support the virus in overcoming innate immune responses by ensuring proper RNA processing and replication. Through these pathways nsp15 contributes significantly to the viral survival and proliferation within the host.

Associated diseases and disorders

Nsp15 is primarily linked to COVID-19. Its role in immune evasion makes it a target of interest in the study of viral pathogenesis and therapeutic interventions. Research has indicated potential interactions between nsp15 and host proteins that could impact disease progression. Furthermore its manipulation or inhibition could serve as a therapeutic strategy potentially reducing the ability of SARS-CoV-2 to spread within the host. Understanding these interactions could aid in the development of antiviral drugs targeting nsp15 specifically.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

8 product images

  • Western blot - Anti-SARS-CoV-2 nsp15 antibody [EPR24840-74] (ab284629), expandable thumbnail

    Western blot - Anti-SARS-CoV-2 nsp15 antibody [EPR24840-74] (ab284629)

    ab284629 was shown to bind specifically to SARS-CoV-2 NSP15 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.

    Lanes 1 - 2: Western blot - Anti-SARS-CoV-2 nsp15 antibody [EPR24840-74] (ab284629)

    Lanes 3 - 4: Western blot - Anti-SARS-CoV-2 nsp15 antibody [EPR24840-74] (ab284629) at 1/1000 dilution

    Lanes 1 and 3: Recombinant SARS-CoV2 NSP15 protein (His tagged) at 0.1 µg

    Lanes 2 and 4: Recombinant SARS-CoV2 NSP15 protein (His tagged) at 0.05 µg

    Secondary

    Lanes 1 - 2: Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution

    Lanes 3 - 4: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/20000 dilution

    Predicted band size: 40 kDa

    Observed band size: 40 kDa

  • Indirect ELISA - Anti-SARS-CoV-2 nsp15 antibody [EPR24840-74] (ab284629), expandable thumbnail

    Indirect ELISA - Anti-SARS-CoV-2 nsp15 antibody [EPR24840-74] (ab284629)

    Indirect ELISA showing primary antibody ab284629 binding to recombinant SARS-COV-2 NSP15 His-tag protein. Plates were coated with recombinant SARS-COV-2 NSP15 His-tag protein and recombinant SARS-COV-2 NSP9 His-tag protein at 1000 ng/ml. Binding of ab284629 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, Goat Anti-Rabbit IgG H&L (HRP) preadsorbed ab97080) at 1/2000 dilution.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SARS-CoV-2 nsp15 antibody [EPR24840-74] (ab284629), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SARS-CoV-2 nsp15 antibody [EPR24840-74] (ab284629)

    Immunohistochemical analysis of Paraffin-embedded sections of mouse spleen tissue labelling SARS-CoV-2 NSP15 with ab284629 at 1/100 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Counter stained with Haematoxylin. Secondary antibody only control: Used PBS instead of primary antibody

    Negative control: no staining on mouse spleen.

    The section was incubated with ab284629 for 30 mins at room temperature. Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 1 (pH 6.0) for 20 minutes.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SARS-CoV-2 nsp15 antibody [EPR24840-74] (ab284629), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SARS-CoV-2 nsp15 antibody [EPR24840-74] (ab284629)

    Immunohistochemical analysis of Paraffin-embedded sections of rat spleen tissue labelling SARS-CoV-2 NSP15 with ab284629 at 1/100 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Counter stained with Haematoxylin. Secondary antibody only control: Used PBS instead of primary antibody

    Negative control: no staining on rat spleen.

    The section was incubated with ab284629 for 30 mins at room temperature. Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 1 (pH 6.0) for 20 minutes.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SARS-CoV-2 nsp15 antibody [EPR24840-74] (ab284629), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SARS-CoV-2 nsp15 antibody [EPR24840-74] (ab284629)

    Immunohistochemical analysis of Paraffin-embedded sections A) HEK-293T transfected with SARS-CoV2 NSP15 expression vector containing a his tag (B) HEK-293T transfected with SARS-CoV-2 NSP9 expression vector containing a his tag labelling SARS-CoV-2 NSP15 with ab284629 at 1/100 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Counter stained with Haematoxylin. Secondary antibody only control: Used PBS instead of primary antibody

    Positive staining on HEK-293T transfected with SARS-CoV2 NSP15 expression vector containing a his tag (Image A); No staining on HEK-293T SARS-CoV-2 NSP9 expression vector containing a his tag. (Image B).

    The section was incubated with ab284629 for 30 mins at room temperature. Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 1 (pH 6.0) for 20 minutes.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SARS-CoV-2 nsp15 antibody [EPR24840-74] (ab284629), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SARS-CoV-2 nsp15 antibody [EPR24840-74] (ab284629)

    Immunohistochemical analysis of Paraffin-embedded sections of human tonsil tissue labelling SARS-CoV-2 NSP15 with ab284629 at 1/100 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Counter stained with Haematoxylin. Secondary antibody only control: Used PBS instead of primary antibody

    Negative control: no staining on human tonsil.

    The section was incubated with ab284629 for 30 mins at room temperature. Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 1 (pH 6.0) for 20 minutes.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

  • Flow Cytometry - Anti-SARS-CoV-2 nsp15 antibody [EPR24840-74] (ab284629), expandable thumbnail

    Flow Cytometry - Anti-SARS-CoV-2 nsp15 antibody [EPR24840-74] (ab284629)

    Flow Cytometry analysis of HEK-293T (Human embryonic kidney epithelial cell) cells transfected with a SARS-CoV-2 nsp9 (Left) or SARS-CoV2 nsp15 (Right) expression vector containing a myc tag labeling SARS-CoV2 nsp15 with ab284629 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® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) secondary antibody was used at 1/5000 dilution. Alexa Fluor® 647 Anti-Myc tag antibody [9E10] (Alexa Fluor® 647 Anti-Myc tag antibody [9E10] ab223895) was used to label transfected cells.

  • Immunocytochemistry/ Immunofluorescence - Anti-SARS-CoV-2 nsp15 antibody [EPR24840-74] (ab284629), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-SARS-CoV-2 nsp15 antibody [EPR24840-74] (ab284629)

    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-CoV2 nsp15 with primary antibody anti-SARS-CoV-2 nsp15 antibody (ab284629) at 1/500 dilution, followed by AlexaFluor®488 Goat anti-Rabbit secondary antibody (green) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/1000 dilution. Myc-Tag Mouse mab (Alexa Fluor® 647) (red) was used to counterstain at 1/100 dilution. The nuclear counter stain is DAPI (blue).

    Confocal image showing positive staining in HEK-293T cells transfected with a SARS-CoV2 nsp15 expression vector containing a myc tag, and no staining in HEK-293T cells transfected with a SARS-CoV2 nsp9 expression vector containing a myc tag. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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Product protocols

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