Anti-Protein Kinase D2 antibody

• Supplier: Abcam
• Catalog: ab7281
• Prices: $400.00
• Sizes: 50 µg
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Applications: WB IHC-P ICC/IF ICC/IF IP
• Species Reactivities: Mouse, Human
• Antigen: Human protein kinase D2
• Description: Rabbit Polyclonal
• Gene: PRKD2
• Conjugate: Unconjugated

Product images

ab7281 staining Protein Kinase D2 in human BON cells (derived from a human pancreatic carcinoid tumor) by Immunocytochemistry/ Immunofluorescence.BON cells were grown on glass coverslips in 24-well plates. 2 or 3 days after seeding or transfection, cells were treated with vehicle (Me2SO) or PMA (100 nm) for 30 minutes. After treatment, cells were fixed with 4% paraformaldehyde for 20 minutes at 37 °C. After three washes with PBS, the cells were permeabilized with 0.3% Triton X-100 for 15 minutes at 37 °C and blocked with 1% bovine serum albumin/PBS for 10 minutes. The cells were incubated with the primary antibody diluted with 1% bovine serum albumin/PBS for 1 hour at room temperature. Cells were washed three times with PBS and incubated with Alexa-conjugated secondary antibody diluted by 1/500 in 1% bovine serum albumin/PBS. Double staining was performed using antibodies ab7281 and an anti-NT. Co-localization is shown by arrows in the merged images.

Anti-Protein Kinase D2 antibody (ab7281) at 1/500 dilution + whole cell lysate prepared from murine NIH3T3 cells at 25 µgSecondaryDonkey anti-rabbit HRP at 1/5000 dilutiondeveloped using the ECL techniqueExposure time : 15 minutesImage courtesy of an anonymous Abreview.

ICC/IF image of ab7281 stained HeLa cells. The cells were 4% formaldehyde (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab7281, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

Protein Kinase D2 was immunoprecipitated using 0.5mg NIH3T3 whole cell extract, 5µg of Rabbit polyclonal to Protein Kinase D2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, NIH3T3 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab7281.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 101KDa: Protein Kinase D2

Product References

Ultraviolet B irradiation and activation of protein kinase D in primary mouse - Ultraviolet B irradiation and activation of protein kinase D in primary mouse

Arun SN, Kaddour-Djebbar I, Shapiro BA, Bollag WB. Oncogene. 2011 Mar 31;30(13):1586-96.

PKD1, PKD2, and their substrate Kidins220 regulate neurotensin secretion in the - PKD1, PKD2, and their substrate Kidins220 regulate neurotensin secretion in the

Li J, Chen LA, Townsend CM Jr, Evers BM. J Biol Chem. 2008 Feb 1;283(5):2614-21. Epub 2007 Nov 29.

Overexpression of wild-type PKD2 leads to increased proliferation and invasion of - Overexpression of wild-type PKD2 leads to increased proliferation and invasion of

Jackson LN, Li J, Chen LA, Townsend CM, Evers BM. Biochem Biophys Res Commun. 2006 Sep 29;348(3):945-9. Epub 2006 Jul 31.