Anti-VDAC1/Porin + VDAC2 + VDAC3 antibody – Mitochondrial Loading Control

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VDAC1/Porin + VDAC2 + VDAC3 antibody – Mitochondrial Loading Control (AB15895)

IHC image of VDAC1/Porin + VDAC2 + VDAC3 staining in a section of formalin-fixed paraffin-embedded normal human heart performed on a Leica Biosystems BOND® RX instrument. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab15895, 0.1 μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-VDAC1/Porin + VDAC2 + VDAC3 antibody – Mitochondrial Loading Control (AB15895)

ab15895 staining VDAC1/Porin + VDAC2 + VDAC3 in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab15895 at 5µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VDAC1/Porin + VDAC2 + VDAC3 antibody – Mitochondrial Loading Control (AB15895)

Immunohistochemical analysis of formalin fixed paraffin embedded human heart labelling VDAC1/Porin with ab15895 at a concentration of 0.1µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

ab15895 Anti-VDAC1/Porin antibody was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)

• WB

Unknown

Western blot - Anti-VDAC1/Porin + VDAC2 + VDAC3 antibody – Mitochondrial Loading Control (AB15895)

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.

All lanes:

Western blot - Anti-VDAC1/Porin + VDAC2 + VDAC3 antibody – Mitochondrial Loading Control (ab15895)

Predicted band size: 31 kDa

true

Exposure time: 3min

• WB

Project

Western blot - Anti-VDAC1/Porin + VDAC2 + VDAC3 antibody – Mitochondrial Loading Control (AB15895)

All lanes:

Western blot - Anti-VDAC1/Porin + VDAC2 + VDAC3 antibody – Mitochondrial Loading Control (ab15895) at 1 µg/mL

Lane 1:

Hela Nuclear lysate at 20 µg

Lane 2:

Hela cell lysate at 20 µg

Lane 3:

A431 cell lysate at 20 µg

Lane 4:

Jurkat cell lysate at 20 µg

Lane 5:

HEK293 cell lysate at 20 µg

Lane 6:

Hela Nuclear lysate at 20 µg with Human VDAC1/Porin peptide (<a href='/en-us/products/unavailable/human-vdac1porin-peptide-ab16131'>ab16131</a>)

Lane 7:

Hela cell lysate at 20 µg with Human VDAC1/Porin peptide (<a href='/en-us/products/unavailable/human-vdac1porin-peptide-ab16131'>ab16131</a>)

Lane 8:

A431 cell lysate at 20 µg with Human VDAC1/Porin peptide (<a href='/en-us/products/unavailable/human-vdac1porin-peptide-ab16131'>ab16131</a>)

Lane 9:

Jurkat cell lysate at 20 µg with Human VDAC1/Porin peptide (<a href='/en-us/products/unavailable/human-vdac1porin-peptide-ab16131'>ab16131</a>)

Lane 10:

HEK293 cell lysate at 20 µg with Human VDAC1/Porin peptide (<a href='/en-us/products/unavailable/human-vdac1porin-peptide-ab16131'>ab16131</a>)

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab6721'>ab6721</a>) at 1/5000 dilution

Predicted band size: 31 kDa

Observed band size: 31 kDa

true

• WB

Project

Western blot - Anti-VDAC1/Porin + VDAC2 + VDAC3 antibody – Mitochondrial Loading Control (AB15895)

All lanes:

Western blot - Anti-VDAC1/Porin + VDAC2 + VDAC3 antibody – Mitochondrial Loading Control (ab15895) at 1 µg/mL

Lane 1:

Chicken liver cell lysate at 20 µg

Lane 2:

CHO K1 cell lysate at 20 µg

Lane 3:

MDCK cell lysate at 20 µg

Lane 4:

Chicken liver cell lysate at 20 µg with Human VDAC1/Porin peptide (<a href='/en-us/products/unavailable/human-vdac1porin-peptide-ab16131'>ab16131</a>)

Lane 5:

CHO K1 cell lysate at 20 µg with Human VDAC1/Porin peptide (<a href='/en-us/products/unavailable/human-vdac1porin-peptide-ab16131'>ab16131</a>)

Lane 6:

MDCK cell lysate at 20 µg with Human VDAC1/Porin peptide (<a href='/en-us/products/unavailable/human-vdac1porin-peptide-ab16131'>ab16131</a>)

Secondary

All lanes:

Alexa fluor goat polyclonal anti-Rabbit IgG at 1/10000 dilution

Predicted band size: 31 kDa

Observed band size: 31 kDa

true

• WB

Project

Western blot - Anti-VDAC1/Porin + VDAC2 + VDAC3 antibody – Mitochondrial Loading Control (AB15895)

All lanes:

Western blot - Anti-VDAC1/Porin + VDAC2 + VDAC3 antibody – Mitochondrial Loading Control (ab15895) at 1 µg/mL

Lane 1:

Heart (Mouse) Tissue Lysate at 10 µg

Lane 2:

Kidney (Mouse) Tissue Lysate at 10 µg

Lane 3:

Western blot - Mouse skeletal muscle tissue lysate - total protein (<a href='/en-us/products/tissue-lysates/mouse-skeletal-muscle-tissue-lysate-total-protein-ab29711'>ab29711</a>) at 10 µg

Lane 4:

Spinal Cord (Mouse) Tissue Lysate at 10 µg

Lane 5:

PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 10 µg

Lane 6:

Brain (Rat) Tissue Lysate - normal tissue at 10 µg

Lane 7:

Kidney (Rat) Whole Cell Lysate - normal tissue (<a href='/en-us/products/unavailable/kidney-rat-whole-cell-lysate-normal-tissue-ab29480'>ab29480</a>) at 10 µg

Secondary

All lanes:

IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Predicted band size: 31 kDa

Observed band size: 31 kDa

false