Anti-VDAC1 / Porin antibody - Mitochondrial Loading Control (ab15895)

Paraffin embedded sections of human kidney were incubated with ab15895 (1/600 dilution) at room temperature for 30 mins. Antigen retrieval was performed by heat induction in citrate buffer pH 6. ab15895 was also tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further images can be found at www.proteinatlas.org

ab15895 stained in HeLa cells. Cells were fixed with 100% methanol (5 min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab15895 at 5µg/ml and ab7291 (Mouse monoclonal to alpha Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed, (pseudo-colored green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594) preadsorbed, (colored red), both used at a 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1hour at room temperature.

ab15895 staining VDAC1/Porin in Human Testis tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 2% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in citric acid. Samples were incubated with primary antibody (1/50 in TBS/BSA/azide buffer) for 16 hours at 21°C. A Biotin-conjugated Goat anti-rabbit IgG polyclonal (1/300) was used as the secondary antibody.

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ab15895 staining mouse heart tissue section by IHC-P.  Sections were PFA fixed and subjected to heat mediated antigen retrieval in citrate buffer (pH 6) prior to blocking in 10% serum for 10 minutes at 25°C.  The primary antibody was diluted 1/600 and incubated with the sample for 1 hour at 25°C.  A biotinylated goat anti-rabbit antibody diluted 1/400 was used as the secondary.

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Western blot image using the Optiblot Reducing Electrophoresis Kit - 10 x 10 cm (4-20%) (ab119220) with the
Prism Ultra Protein Ladder (ab116028) 5µl used. We recommend using our ECL substrate kit (ab65623) .
20ug of Lysate per lane and detection using ab15895 diluted to 1ug/ml.
Lane 1: HeLa cell lysate
Lane 2: Jurkat cell lysate
Lane 3: A431 cell lysate
Lane 4: HEK293 cell lysate
Lane 5: HepG2 cell lysate.