Anti-VDAC1/Porin antibody [EPR27552-6] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VDAC1/Porin antibody [EPR27552-6] - BSA and Azide free (AB306582)

This data was developed using ab306581, the same antibody clone in a different buffer formulation. 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] - BSA and Azide free (AB306582)

This data was developed using ab306581, the same antibody clone in a different buffer formulation. 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] - BSA and Azide free (AB306582)

This data was developed using ab306581, the same antibody clone in a different buffer formulation. 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] - BSA and Azide free (AB306582)

This data was developed using 306581, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : The 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 ЎгC. 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. Exposure time :

All lanes:

Western blot - Anti-VDAC1/Porin antibody [EPR27552-6] - Mitochondrial Loading Control (<a href='/en-us/products/primary-antibodies/vdac1-porin-antibody-epr27552-6-mitochondrial-loading-control-ab306581'>ab306581</a>) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate 20 μg

Lane 2:

VDAC1/Porin knockout HAP1 whole cell lysate 20 μg

Lane 3:

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

Lane 4:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate 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] - BSA and Azide free (AB306582)

This data was developed using ab306581, the same antibody clone in a different buffer formulation. 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] - BSA and Azide free (AB306582)

This data was developed using 306581, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : Low expression tissue : liver (PMID : 31167495). The identity of the higher MW band at approximately 160 kDa is unknown. 180 seconds Exposure time :

All lanes:

Western blot - Anti-VDAC1/Porin antibody [EPR27552-6] - Mitochondrial Loading Control (<a href='/en-us/products/primary-antibodies/vdac1-porin-antibody-epr27552-6-mitochondrial-loading-control-ab306581'>ab306581</a>) 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