- MA5-24964 - Invitrogen Antibodies POR antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of POR in HEK293T cells in untransfected (Left lane) and transfected (Right lane) samples using 5 µg per lane. The samples were separated by SDS-PAGE and probed with POR (Product # MA5-24964) monoclonal antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of POR in HEK293T cells in untransfected (Left lane) and transfected (Right lane) samples using 5 µg per lane. The samples were separated by SDS-PAGE and probed with POR (Product # MA5-24964) monoclonal antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of POR in HepG2, HeLa, SVT2, A549, COS7, Jurkat, MDCK, PC12 and MCF7 cells using 35 µg per lane. Samples were separated by SDS-PAGE and probed with POR (Product # MA5-24964) monoclonal antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of NADPH--cytochrome P450 reductase was achieved by transfecting HeLa with NADPH--cytochrome P450 reductase specific siRNAs (Silencer® select Product # S10839, S10838). Western Blot analysis (Fig. a) was performed using Whole cell extracts from the NADPH--cytochrome P450 reductase knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The Blot was probed with Cytochrome P450 Reductase Monoclonal Antibody (OTI6C4) (Product # MA5-24964, 1:2000 dilution ) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution). Densitometric analysis of this western Blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to NADPH--cytochrome P450 reductase.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot was performed using Anti-Cytochrome P450 Reductase Monoclonal Antibody (OTI6C4) (Product # MA5-24964) and a 77 kDa band corresponding to NADPH-cytochrome P450 reductase was observed across the cell lines and tissues tested except Rat Liver. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), Hep G2 (Lane 2), NIH:OVCAR-3 (Lane 3), Mouse Kidney (Lane 4), Rat Liver (Lane 5) and Mouse Liver (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2001) by iBlot® 2 Dry Blotting System (Product # IB21001). The Blot was probed with the primary antibody (1:2000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of POR in COS7 cells. Cells were transfected with a plasmid overexpressing POR and probed with a POR monoclonal antibody (Product # MA5-24964).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of POR in COS7 cells. Cells were transfected with a plasmid overexpressing POR and probed with a POR monoclonal antibody (Product # MA5-24964).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of NADPH-cytochrome P450 reductase was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Cytochrome P450 Reductase Monoclonal Antibody (OTI6C4) (Product # MA5-24964) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjµgate (Product # A28175), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing endoplasmic reticulum like localization. Panel e represents control HeLa cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry was performed on paraffin-embedded human endometrium tissue. To expose target proteins, 10mM citric buffer, pH6.0, 100°C for 10min was used. Following antigen retrieval, tissues were probed with a POR monoclonal antibody (Product # MA5-24964).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry was performed on paraffin-embedded human ovary tissue. To expose target proteins, 10mM citric buffer, pH6.0, 100°C for 10min was used. Following antigen retrieval, tissues were probed with a POR monoclonal antibody (Product # MA5-24964).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry was performed on paraffin-embedded human colon tissue. To expose target proteins, 10mM citric buffer, pH6.0, 100°C for 10min was used. Following antigen retrieval, tissues were probed with a POR monoclonal antibody (Product # MA5-24964).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on paraffin-embedded human breast tissue. To expose target proteins, 10mM citric buffer, pH6.0, 100°C for 10min was used. Following antigen retrieval, tissues were probed with a POR monoclonal antibody (Product # MA5-24964).

MA5-24964