PA5-59539

antibody from Invitrogen Antibodies

Targeting: OAS3

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-59539

Provider

Invitrogen Antibodies

Product name

OAS3 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant full-length protein

Description

Immunogen sequence: TWDLGNGAAW HWDLLAQEAA SCYDHPCFLR GMGDPVQSWK GPGLPRAGCS GLGHPIQLDP NQKTPENSKS LNAVYPRAGS KPPSCP

Concentration

0.4 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of OAS3 in Lane 1: Marker (kDa) 250, 130, 95, 72, 55, 36, 28, 17, 10; Lane 2: Human cell line RT-4. Samples were probed using an OAS3 Polyclonal Antibody (Product # PA5-59539).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot was performed using Anti-OAS3 Polyclonal Antibody (Product # PA5-59539) and a 120 kDa band corresponding to 2-5-oligoadenylate synthase 3 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), PC-3 (Lane 7) and DU 145 (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 (0.2 µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 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 OAS3 was observed upon Interferon treatment in all the models tested (https://doi.org/10.1073/pnas.1519657113).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent staining of OAS3 in human cell line U-251 MG shows positivity in cytoplasm & nucleus but excluded from the nucleoli. Samples were probed using an OAS3 Polyclonal Antibody (Product # PA5-59539).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of 2-5-oligoadenylate synthase 3 was performed using 70% confluent log phase A549 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 OAS3 Polyclonal Antibody (Product # PA5-59539) at 1 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (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 predominant 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).

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical staining of OAS3 in human kidney tissue shows strong cytoplasmic and membranous positivity in cells in tubules. Samples were probed using an OAS3 Polyclonal Antibody (Product # PA5-59539).