ATG12 Antibody PA5-17217 from Thermo Fisher Scientific

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WB

Western blot was performed using Anti-ATG12 Polyclonal Antibody(Product # PA5-17217) and a 17kDa band corresponding to ATG12 was observed upon MG132 treatment in U-2OS. A 55 kDa band corresponding to the covalently conjugated ATG12-ATG5 was observed across cell lines tested. Whole Cell Extract-TEB (30 µg lysate) of HCT 116 (Lane 1), U-2 OS (Lane 2), U-2OS treated with MG132 (10uM for 16 hr) (Lane 3), HeLa (Lane 4) and Hep G2 (Lane 5) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0342BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).
ICC/IF

Immunofluorescence analysis of ATG12 was performed using 70% confluent log phase HCT 116 cells. The cells were fixed with 4% paraformaldehyde for 10 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 ATG12 Polyclonal Antibody (Product # PA5-17217) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731), (1:2000), 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). Panel d represents the merged image showing cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
ATG12 Antibody

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Thermo Fisher Scientific for
ATG12 Antibody

Thermo Fisher Scientific's ATG12 Antibody is a Rabbit Polyclonal antibody. This antibody has been shown to work in applications such as: Immunocytochemistry, and Western Blot. The ATG12 Antibody was generated using ATG12 as the antigen and it reacts with Homo Sapiens, and Human.

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ICC/IF

ATG12 Antibody

ATG12 Antibody from Thermo Fisher Scientific

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ATG12 Antibody from Thermo Fisher Scientific

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