- 34-8900 - Invitrogen Antibodies CDC14B antibody

Cho HP, Liu Y, Gomez M, Dunlap J, Tyers M, Wang Y
Molecular and cellular biology 2005 Jun;25(11):4541-51

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts and tissue lysate of Hep G2 (Lane 1), U-2 OS (Lane 2), HeLa (Lane 3) and Mouse Cerebellum (Lane 4). The blots were probed with Anti- CDC14B Rabbit Polyclonal Antibody (Product # 34-8900, 2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conj µgate (Product # A27036, 0.4 µg/mL, 1:2500 dilution. Bands of ~ 53 and 55 kDa corresponding to CDC14B were observed across cell lines and tissue lysate tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts and tissue lysate of Hep G2 (Lane 1), U-2 OS (Lane 2), HeLa (Lane 3) and Mouse Cerebellum (Lane 4). The blots were probed with Anti- CDC14B Rabbit Polyclonal Antibody (Product # 34-8900, 2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conj µgate (Product # A27036, 0.4 µg/mL, 1:2500 dilution. Bands of ~ 53 and 55 kDa corresponding to CDC14B were observed across cell lines and tissue lysate tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent staining of U2OS or HeLa cells with Rb anti-CDC14B (green) (Product # 34-8900). DNA is labeled with DAPI (blue), anti-CDC14B is labeled with FITC (green), and anti-Tubulin is labeled with rhodamine (red). CDC14B appears as bright nuclear dots and midbodies of cells undergoing cytokinesis. Image courtesy of Dr. Catherine Regnard and Dr. Ulrike Grueneberg, Max Planck Institute for Biochemistry, Germany.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of Cdc14B was done on Hep G2 cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with Cdc14B Rabbit Polyclonal Antibody (Product # 34-8900, red histogram) or with rabbit isotype control (pink histogram) at 3-5 µg/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Goat Anti-Rabbit Secondary Antibody (Product # A11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10, 000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.

34-8900