- PA5-61433 - Invitrogen Antibodies XRN2 antibody

No submitted references: Submit reference

No comments: Submit comment

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of XRN2 in Lane 1: Marker (kDa) 250, 130, 95, 72, 55, 36, 28, 17, 10; Lane 2: Human cell line RT-4; Lane 3: Human cell line U-251MG sp. Samples were probed using a XRN2 Polyclonal Antibody (Product # PA5-61433).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of XRN2 was achieved by transfecting HeLa with XRN2 specific siRNAs (Silencer® select Products # s22413, s22412). Western blot analysis (Fig. a) was performed using whole cell extracts from the XRN2 knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with XRN2 Polyclonal Antibody (Product # PA5-61433, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to XRN2.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-XRN2 Polyclonal Antibody (Product # PA5-61433) and a 110kDa band corresponding to XRN2 was observed in all the tested cell models along with some uncharacterized bands at ~80-90 kDa. Modified whole cell lysates (1% SDS) (30ug lysate) of HEK-293 (Lane 1), HeLa (Lane 2), COS-7 (Lane 3), THP-1 (Lane 4) and MCF7 (Lane 5) were electrophoresed using Novex® NuPAGE® 10% Bis-Tris gel (Product # NP0302BOX). 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 (1:1000 dilution) and detected by chemiluminescence with Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent staining of XRN2 in human cell line U-2 OS shows positivity in nucleus & nucleoli. Samples were probed using a XRN2 Polyclonal Antibody (Product # PA5-61433).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of XRN2 was performed using HeLa cells. 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 XRN2 Polyclonal Antibody (Product # PA5-61433) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 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 nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical staining of XRN2 in human colon tissue shows strong nuclear and cytoplasmic positivity in glandular cells. Samples were probed using a XRN2 Polyclonal Antibody (Product # PA5-61433).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Chromatin Immunoprecipitation (ChIP) assay of endogenous XRN2 protein using Anti-XRN2 Antibody: ChIP was performed using Anti-XRN2 Rabbit Polyclonal Antibody (Product # PA5-61433, 5 µg) on sheared chromatin from HeLa cells using the MAGnify ChIP System kit (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR using primers binding to MALAT2 transcriptional start site and SAT2 satellite repeats. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

PA5-61433