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Synthetic peptide derived from residues 150 - 250 of Human VDAC1/ Porin. The immunogen sequence is completely conserved between isoforms 1, 2 and 3 of human VDAC (Note: the amino acid sequence is proprietary; Peptide available as ab16131.)
Our Abpromise guarantee covers the use of ab15895 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 31 kDa (predicted molecular weight: 31 kDa).
Abcam recommends using BSA as the blocking agent.
|IHC-P||Use at an assay dependent concentration.|
|ICC/IF||Use a concentration of 5 µg/ml.|
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
ICC/IF image of ab15895 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab15895, 5µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 phalloidin was used to label F-actin (red).
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.
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.
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.