Anti-VDAC1/Porin antibody [EPR27552-6] - Mitochondrial Loading Control

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VDAC1/Porin antibody [EPR27552-6] - Mitochondrial Loading Control (AB306581)

Immunohistochemical analysis of paraffin-embedded Human liver tissue lABeling VDAC1/Porin with ab306581 at 1/5000 (0.104 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining in human liver (PMID : 31167495). The section was incubated with ab306581 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VDAC1/Porin antibody [EPR27552-6] - Mitochondrial Loading Control (AB306581)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue lABeling VDAC1/Porin with ab306581 at 1/5000 (0.104 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining in rat kidney. The section was incubated with ab306581 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VDAC1/Porin antibody [EPR27552-6] - Mitochondrial Loading Control (AB306581)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue lABeling VDAC1/Porin with ab306581 at 1/5000 (0.104 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining in mouse kidney (PMID : 27907123). The section was incubated with ab306581 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• WB

Supplier Data

Western blot - Anti-VDAC1/Porin antibody [EPR27552-6] - Mitochondrial Loading Control (AB306581)

Blocking and diluting buffer and concentration : Intercept^®(TBS) Blocking Buffer diluted with an equal volume of 0.1% TBSThe samples were run on a Bis-Tris gel.
Performed under reducing conditions.
False colour image of Western blot : Anti-VDAC1/Porin antibody [EPR27552-6] (ab306581) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab306581 was shown to bind specifically to VDAC1/Porin. A band was observed at 33 kDa in wild-type HAP1 cell lysates whereas no signal observed at this size in VDAC1/Porin knockout cell line. To generate this image, wild-type and VDAC1/Porin knockout HAP1 cell lysates were analyzed. First, samples were run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in in Intercept^® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-VDAC1/Porin antibody [EPR27552-6] - Mitochondrial Loading Control (ab306581) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

VDAC1/Porin knockout HAP1 whole cell lysate at 20 µg

Lane 3:

HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 20 µg

Secondary

Lanes 1 - 4:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Lanes 1 - 4:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution

Observed band size: 33 kDa

false

• Dot

Supplier Data

Dot Blot - Anti-VDAC1/Porin antibody [EPR27552-6] - Mitochondrial Loading Control (AB306581)

Dot blot analysis of VDAC1/Porin using ab306581 at 1 : 1000 (0.518 ug/ml) followed by a Goat Anti-RABbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution. Exposure time : 180 seconds Blocking and diluting buffer and concentration : 5% NFDM/TBST

• WB

Supplier Data

Western blot - Anti-VDAC1/Porin antibody [EPR27552-6] - Mitochondrial Loading Control (AB306581)

Blocking and diluting buffer and concentration : 5% NFDM/TBSTLow expression tissue : liver (PMID : 31167495). The identity of the higher MW band at approximately 160 kDa is unknown. Exposure time : 180 seconds

All lanes:

Western blot - Anti-VDAC1/Porin antibody [EPR27552-6] - Mitochondrial Loading Control (ab306581) at 1/1000 dilution

Lane 1:

Mouse kidney tissue lysate 20 μg

Lane 2:

Mouse liver tissue lysate 20 μg

Lane 3:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 20 μg

Lane 4:

C6 (rat glial tumor glial cell) whole cell lysate 20 μg

Lane 5:

PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate 20 μg

Lane 6:

RAW 264.7 (mouse elson murine leukemia virus-induced tumor macrophage) whole cell lysate 20 μg

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-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 33 kDa

false

Exposure time: 180s