- PA5-29781 - Invitrogen Antibodies BUB3 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: BUB3 antibody detects BUB3 protein by western blot analysis. Various whole cell extracts (30 µg) were separated by 10% SDS-PAGE, and the membrane was blotted with BUB3 antibody Bub3 Polyclonal Antibody (Product # PA5-29781) diluted by 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of Bub3 was performed by separating 30 µg of various whole cell extracts by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a Bub3 Polyclonal Antibody (Product # PA5-29781) at a dilution of 1:500 and a HRP-conjugated anti-rabbit IgG secondary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using Bub3 Polyclonal Antibody (Product # PA5-29781). Sample (30 µg of whole cell lysate). Lane A: 293T. Lane B: H1299. 10% SDS PAGE. Bub3 Polyclonal Antibody (Product # PA5-29781) diluted at 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: BUB3 antibody detects BUB3 protein by western blot analysis. Various whole cell extracts (30 µg) were separated by 10% SDS-PAGE, and the membrane was blotted with BUB3 antibody Bub3 Polyclonal Antibody (Product # PA5-29781) diluted by 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of Bub3 was performed by separating 30 µg of various whole cell extracts by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a Bub3 Polyclonal Antibody (Product # PA5-29781) at a dilution of 1:500 and a HRP-conjugated anti-rabbit IgG secondary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: KD of Bub3 was achieved by transfecting HeLa with Bub3 specific siRNAs (Silencer® select Product # s17556, s2451). Western blot analysis (Fig. a) was performed using modified whole cell extracts (1% SDS) from the Bub3 KD cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with Bub3 Polyclonal Antibody (Product # PA5-29781, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 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 Bub3..

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-Bub3 Polyclonal Antibody (Product # PA5-29781) and 37 kDa band corresponding to Bub3 was observed across cell lines and tissues tested. Modified whole cell extracts (1% SDS) (30 µg lysate) of HeLa (Lane 1), A549 (Lane 2), A-431 (Lane 3), Hep G2 (Lane 4), PC-3 (Lane 5), HEK-293 (Lane 6), Jurkat (Lane 7), SK-BR-3 (Lane 8), tissue extracts of Mouse Brain (Lane 9) and Mouse Spleen (Lane 10) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris 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 (1:1000 dilution) 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: Immunofluorescence analysis of Bub3 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 1 hour at room temperature. The cells were labeled with Bub3 Polyclonal Antibody (Product # PA5-29781) at 1:100 dilution 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 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 analysis of paraffin-embedded A549 Xenograft, using BUB3 (Product # PA5-29781) antibody at 1:100 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunoprecipitation of BUB3 was performed in 293T whole cell extracts using 5 µg of Bub3 Polyclonal Antibody (Product # PA5-29781). Samples were transferred to a membrane and probed with Bub3 Polyclonal Antibody as a primary antibody and an HRP-conjugated anti-Rabbit IgG was used as a secondary antibody.

PA5-29781