Invitrogen

OAS2 Monoclonal Antibody (OTI5E1)

Advanced Verification

This Antibody was verified by Cell treatment to ensure that the antibody binds to the antigen stated.

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Datasheet

Protocols

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OAS2 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of 2-5-oligoadenylate synthase 2 was performed using 70% confluent log phase A549 cells treated with IFN beta 1A (1000 U/mL, O/N). The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with OAS2 Monoclonal Antibody (OTI5E1) (Product # MA5-26557) at 1:200 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing predominantly Cytoplasmic localization. Panel e represents untreated A549 cells. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification in EVOS™ M7000 Imaging System (Product # AMF7000).

OAS2 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry was performed on paraffin-embedded human lymph node tissue. To expose target proteins, 10mM citric buffer, pH6.0, 120°C for 3min was used. Following antigen retrieval, tissues were probed with a OAS2 monoclonal antibody (Product # MA5-26557).

OAS2 Antibody in Western Blot (WB) — Western Blot was performed using Anti-OAS2 Monoclonal Antibody (OTI5E1) (Product # MA5-26557) and a 70 kDa band corresponding to 2-5-oligoadenylate synthase 2 was observed. Whole cell extracts (30 µg lysate) of A549 (Lane 1), A549 treated with IFN beta 1A (1000 U/mL, O/N) (Lane 2), HT-1080 (Lane 3), HT-1080 treated with IFN alpha A (2000 U/mL, O/N) (Lane 4), HeLa (Lane 5), HeLa treated with IFN alpha A (10 ng/mL,16 hrs) (Lane 6), Jurkat (Lane 7) and HEK-293 (Lane 8) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:2000) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Upregulation in the expression of OAS2 was observed upon Interferon treatment in all the models tested (https://doi.org/10.1073/pnas.1519657113). Being a very low expresser of OAS2, band of interest was not observed in HEK-293 model as expected.

OAS2 Antibody — Altered expression of proteins upon cell treatment demonstrates antibody specificity. Western blot using OAS2 Monoclonal Antibody (OTI5E1) (Product # MA5-26557), shows upregulation in OAS2 expression upon IFN beta 1A treatment in A549, and IFN alpha A treatment in HT-1080 and HeLa (https://doi.org/10.1073/pnas.1519657113). {TM}

Product Details

MA5-26557

Applications	Tested Dilution	Publications
Western Blot (WB)	1:2,000	-
Immunohistochemistry (Paraffin) (IHC (P))	1:150	-
Immunocytochemistry (ICC/IF)	1:200	-

Product Specifications
Species Reactivity	Human
Host/Isotype	Mouse / IgG2a
Class	Monoclonal
Type	Antibody
Clone	OTI5E1
Immunogen	Human recombinant protein fragment corresponding to amino acids 60-335 of OAS2 produced in E.coli View immunogen
Conjugate	Unconjugated
Form	Liquid
Concentration	1 mg/mL
Purification	Affinity Chromatography
Storage buffer	PBS, pH 7.3, with 1% BSA, 50% glycerol
Contains	0.02% sodium azide
Storage conditions	-20° C, Avoid Freeze/Thaw Cycles
RRID	AB_2724897

Target Information

This gene encodes a member of the 2-5A synthetase family, essential proteins involved in the innate immune response to viral infection. The encoded protein is induced by interferons and uses adenosine triphosphate in 2'-specific nucleotidyl transfer reactions to synthesize 2',5'-oligoadenylates. These molecules activate latent RNase L, which results in viral RNA degradation and the inhibition of viral replication. The three known members of this gene family are located in a cluster on chromosome 12. Alternatively spliced transcript variants encoding different isoforms have been described.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Protein Aliases: (2'-5')oligo(A) synthetase 2; (2-5')oligo(A) synthase 2; 2'-5'-oligoadenylate synthase 2; 2'-5'-oligoadenylate synthetase 2, 69/71kDa; 2-5A synthase 2; A synthetase 2; MGC78578; p69 OAS / p71 OAS; p69OAS / p71OAS

Gene Aliases: OAS2

UniProt ID: (Human) P29728

Entrez Gene ID: (Human) 4939

Molecular Function: nucleotidyltransferase

Function(s)

nucleobase-containing compound metabolic process RNA catabolic process protein glycosylation response to virus protein myristoylation defense response to virus interferon-gamma-mediated signaling pathway type I interferon signaling pathway

Process(es)

2'-5'-oligoadenylate synthetase activity double-stranded RNA binding protein binding ATP binding metal ion binding

OAS2 Monoclonal Antibody (OTI5E1)

Product Details

MA5-26557

Western Blot (WB)

Immunohistochemistry (Paraffin) (IHC (P))

Immunocytochemistry (ICC/IF)

Species Reactivity

Host/Isotype

Class

Type

Clone

Immunogen

Conjugate

Form

Concentration

Purification

Storage buffer

Contains

Storage conditions

RRID

Target Information

References (0)

Bioinformatics

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