Anti-Cytochrome P450 Reductase antibody [EPR14479(B)] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytochrome P450 Reductase antibody [EPR14479(B)] - BSA and Azide free (AB250241)

This data was developed using the same antibody clone in a different buffer formulation (ab180597). Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labelling Cytochrome P450 Reductase with ab180597 at 1/1000 (0.1 μg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).The section was counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins. Postive staining on rat kidney. The section was incubated with ab180597 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytochrome P450 Reductase antibody [EPR14479(B)] - BSA and Azide free (AB250241)

This data was developed using the same antibody clone in a different buffer formulation (ab180597). Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labelling Cytochrome P450 Reductase with ab180597 at 1/1000 (0.1 μg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).The section was counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins. Postive staining on mouse kidney. The section was incubated with ab180597 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• WB

Lab

Western blot - Anti-Cytochrome P450 Reductase antibody [EPR14479(B)] - BSA and Azide free (AB250241)

This data was developed using the same antibody clone in a different buffer formulation (ab180597). Blocking and diluting buffer and concentration : 5% NFDM/TBST. Exposure time : Lane 1-3 : 26 seconds; Lane 4-7 : 75 seconds. The identities of the lower MW bands between 37 and 60kDa are unknown.

All lanes:

Western blot - Anti-Cytochrome P450 Reductase antibody [EPR14479(B)] (<a href='/en-us/products/primary-antibodies/cytochrome-p450-reductase-antibody-epr14479b-ab180597'>ab180597</a>) at 1/1000 dilution

Lane 1:

Rat brain tissue lysate at 20 µg

Lane 2:

Rat heart tissue lysate at 20 µg

Lane 3:

Rat spleen tissue lysate at 20 µg

Lane 4:

Rat kidney tissue lysate at 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

Predicted band size: 76 kDa

Observed band size: 75 kDa

false

• WB

Lab

Western blot - Anti-Cytochrome P450 Reductase antibody [EPR14479(B)] - BSA and Azide free (AB250241)

This data was developed using the same antibody clone in a different buffer formulation (ab180597). Blocking and diluting buffer and concentration : 5% NFDM/TBST. Exposure time : Lane 1-3 : 26 seconds; Lane 4-7 : 75 seconds. The identities of the lower MW bands between 37 and 60kDa are unknown.

All lanes:

Western blot - Anti-Cytochrome P450 Reductase antibody [EPR14479(B)] (<a href='/en-us/products/primary-antibodies/cytochrome-p450-reductase-antibody-epr14479b-ab180597'>ab180597</a>) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse heart tissue lysate at 20 µg

Lane 3:

Mouse spleen tissue lysate at 20 µg

Lane 4:

Mouse kidney tissue lysate at 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

Predicted band size: 76 kDa

Observed band size: 75 kDa

false

• WB

Lab

Western blot - Anti-Cytochrome P450 Reductase antibody [EPR14479(B)] - BSA and Azide free (AB250241)

This data was developed using the same antibody clone in a different buffer formulation (ab180597).

Lanes 1- 4 : Merged signal (red and green). Green - ab180597 observed at 80 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab180597 was shown to react with Cytochrome P450 Reductase in wild-type HCT116 cells in western blot. Loss of signal was observed when knockout cell line ab266889 (knockout cell lysate ab257596) was used. Wild-type HCT116 and POR knockout HCT116 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab180597 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Cytochrome P450 Reductase antibody [EPR14479(B)] (<a href='/en-us/products/primary-antibodies/cytochrome-p450-reductase-antibody-epr14479b-ab180597'>ab180597</a>) at 1/1000 dilution

Lane 1:

Wild-type HCT116 cell lysate at 40 µg

Lane 2:

POR knockout HCT116 cell lysate at 40 µg

Lane 2:

Western blot - Human POR (Cytochrome P450 Reductase) knockout HCT116 cell line (<a href='/en-us/products/cell-lines/human-por-cytochrome-p450-reductase-knockout-hct116-cell-line-ab266889'>ab266889</a>)

Lane 3:

HepG2 cell lysate at 40 µg

Lane 4:

A431 cell lysate at 40 µg

Predicted band size: 76 kDa

Observed band size: 80 kDa

false

• WB

Lab

Western blot - Anti-Cytochrome P450 Reductase antibody [EPR14479(B)] - BSA and Azide free (AB250241)

This data was developed using the same antibody clone in a different buffer formulation (ab180597).

Lanes 1-4 : Merged signal (red and green). Green - ab180597 observed at 75 kDa. Red - loading control ab8245 observed at 37 kDa.

ab180597 Anti-Cytochrome P450 Reductase antibody [EPR14479(B)] was shown to specifically react with Cytochrome P450 Reductase in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264996 (knockout cell lysate ab257595) was used. Wild-type and Cytochrome P450 Reductase knockout samples were subjected to SDS-PAGE. ab180597 and Anti-GAPDH antibody [6C5] - Loading Control?(ab8245) were incubated overnight at 4^°C at 1 in 10000 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Cytochrome P450 Reductase antibody [EPR14479(B)] (<a href='/en-us/products/primary-antibodies/cytochrome-p450-reductase-antibody-epr14479b-ab180597'>ab180597</a>) at 1/10000 dilution

Lane 1:

Wild-type HeLa lysate at 20 µg

Lane 2:

Cytochrome P450 Reductase knockout HeLa lysate at 20 µg

Lane 2:

Western blot - Human POR (Cytochrome P450 Reductase) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-por-cytochrome-p450-reductase-knockout-hela-cell-line-ab264996'>ab264996</a>)

Lane 3:

HepG2 lysate at 20 µg

Lane 4:

A431 lysate at 20 µg

Predicted band size: 76 kDa

false