- PA5-55754 - Invitrogen Antibodies DHX9 antibody

Huan W, Zhang J, Li Y, Zhi K
American journal of physiology. Cell physiology 2019 Aug 1;317(2):C262-C269

No comments: Submit comment

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of DHX9 in human cell line MOLT-4 using a DHX9 Polyclonal Antibody (Product # PA5-55754).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of DHX9 in mouse cell line NIH-3T3 and rat cell line NBT-II using a DHX9 Polyclonal Antibody (Product # PA5-55754).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: KD of DHX9 was achieved by transfecting A-431 with DHX9 specific siRNAs (Silencer® select Product # s4019). Western blot analysis (Fig. a) was performed using whole cell extracts from the DHX9 KD cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with DHX9 Polyclonal Antibody (Product # PA5-55754, 0.2µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25µg/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 DHX9.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using anti-DHX9 Polyclonal Antibody (Product # PA5-55754) and a 149 kDa band corresponding to DHX9 was observed across cell lines tested. Whole cell extracts (30 µg lysate) of A-431 (Lane 1), A549 (Lane 2), HeLa (Lane 3) and NIH/3T3 (Lane 4) were electrophoresed using NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # LC2001) by wet transfer using XCell SureLock™ Mini-Cell and XCell II™ Blot Module (Product # EI0002). The blot was probed with the primary antibody (0.2 µg/mL) and detected by chemiluminescence 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 DHX9 in human cell line A-431 shows positivity in nucleus but excluded from the nucleoli. Samples were probed using a DHX9 Polyclonal Antibody (Product # PA5-55754).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of DHX9 was performed using A-431 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 DHX9 Polyclonal Antibody (Product # PA5-55754) at 4 microgram/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 DHX9 in human cerebral cortex tissue shows strong nuclear positivity in neuronal cells. Samples were probed using a DHX9 Polyclonal Antibody (Product # PA5-55754).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Detection of binding of endogenous DHX9 protein to specific RNA using Anti-DHX9 Antibody: RNA Immunoprecipitation (RIP) was performed using Anti-DHX9 Recombinant Rabbit Polyclonal Antibody (Product # PA5-55754, 5 ug) on whole cell lysate from 2 million HCT 116 cells. Normal Rabbit IgG was used as a negative IP control. RNA purified by RiboPure™ RNA Purification Kit (Product # AM1924) was analyzed by RT-PCR using the Power SYBR® Green RNA-to-CT™ 1-Step Kit (Product # 4389986) with RIP primer pairs over HOXA2, JUND mRNA and 18s rRNA. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

PA5-55754