- PA5-17223 - Invitrogen Antibodies PBX1 antibody

Submitted references The murine Pbx1-d lupus susceptibility allele accelerates mesenchymal stem cell differentiation and impairs their immunosuppressive function.

Lu S, Zeumer L, Sorensen H, Yang H, Ng Y, Yu F, Riva A, Croker B, Wallet S, Morel L
Journal of immunology (Baltimore, Md. : 1950) 2015 Jan 1;194(1):43-55

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Pbx1 in extracts from A431 and HeLa cells using Pbx1 polyclonal antibody (Product # PA5-17223).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of PBX1 was achieved by transfecting HeLa with PBX1 specific siRNAs (Silencer® select Product # s10080, s10079). Western blot analysis (Fig. a) was performed using whole cell extracts 30 ug of PBX1 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with PBX1 Polyclonal Antibody (Product # PA5-17223, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b) and (Fig. c) for both the isoforms. Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to PBX1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-PBX1 Polyclonal Antibody(Product # PA5-17223) and a 40 kDa and 50kDa band corresponding to PBX1 was observed across Cell lines and tissue. Whole Cell Extract-WCL (30 ug µg lysate) of HEL 92.1.7 (Lane 1), IMR-32 (Lane 2), NIH:OVCAR-3 (Lane 3), HeLa (Lane 4), COS-7 (Lane 5), Mouse Lung (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2001) 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 (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (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 analysis of Pbx1 in HeLa cells using a Pbx1 polyclonal antibody (Product # PA5-17223) (green). Actin filaments are labeled with a fluorescent red phalloidin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of PBX1 was performed using 70% confluent log phase 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 45 minutes at room temperature. The cells were labeled with PBX1 Polyclonal Antibody (Product # PA5-17223) at 1:100 dilution 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 dilution), 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 dilution). Panel d represents the merged image showing Nuclear and faint cytosolic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X with Oil immersion magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Chromatin Immunoprecipitation (ChIP) assay of endogenous PBX1 protein using PBX1 Antibody: ChIP was performed using Anti-PBX1 Polyclonal Antibody (Product # PA5-17223, 2.5 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 SPOCK2 and MEOX1 (Active), ACTB and SAT2 satellite repeats (Inactive). Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

PA5-17223

Antibody data