ATG12 Antibody MA5-27801 from Thermo Fisher Scientific

Supplier Images

WB

Western blot was performed using Anti-ATG12 Monoclonal Antibody (GT166)(Product # MA5-27801) 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 has also been observed across cell lines and tissue extracts tested. Whole Cell Extract-TEB (30 µg lysate) of HCT 116 (Lane 1), U-2 OS (Lane 2), U-2 OS treated with MG132 (10uM for 16 hr) (Lane 3), HeLa (Lane 4), Hep G2 (Lane 5), NIH/3T3 (Lane 6), C2C12 (Lane 7) and tissue extracts of Mouse Colon (Lane 8), Mouse Kidney (Lane 9) and Mouse Pancreas (Lane 10) 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-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).
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 Monoclonal Antibody (GT166) (Product # MA5-27801) at 5 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32723), (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.

Supplier Page from
Thermo Fisher Scientific for
ATG12 Antibody

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

WB

ICC/IF

ATG12 Antibody from Thermo Fisher Scientific

Thermo Fisher Scientific

Reviews

ATG12 Antibody from Thermo Fisher Scientific

About Biocompare

Advertise with Us

Specialized Search Tools

Site Map