Detection of Human and Mouse ATG12 by Western Blot.
Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, HCT‑116 human colorectal carcinoma cell line, and C2C12 mouse myoblast cell line. PVDF membrane (left) was probed with 0.1 µg/mL of Mouse Anti-Human ATG12 Monoclonal Antibody (Catalog # MAB6807) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). For additional reference, PVDF membrane (right) was probed with 0.5 µg/mL of Mouse Anti-Human/Mouse/Rat ATG5 Monoclonal Antibody (Catalog # MAB5294) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for the ATG5:ATG12 heterodimer at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
ATG12 in RAW 264.7 Mouse Cell Line.
ATG12 was detected in immersion fixed RAW 264.7 mouse monocyte/macrophage cell line untreated (lower panel) or stimulated with LPS (upper panel) using Mouse Anti-Human/Mouse ATG12 Monoclonal Antibody (Catalog # MAB6807) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to autophagosomes. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Preparation and Storage
Sterile PBS to a final concentration of 0.5 mg/mL.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
ATG12 is a 15 kDa cytosolic protein with a C-terminal ubiquitin-like domain. The conjugation of ATG12 to ATG5 is required for the autophagosome pathway of bulk degradation. The ATG5:ATG12 heterodimer associates noncovalently with an ATG16 multimer to generate autophagosomes. The ATG5:ATG12 conjugate also inhibits type I interferon-induced antiviral responses and promotes the lipidation of ATG8. Alternate splicing of human ATG12 generates an isoform that lacks the ubiquitin-like domain. Human ATG12 shares 89% and 85% aa sequence identitiy with mouse and rat ATG12, respectively.
ATG12 Autophagy Related 12 Homolog
Entrez Gene IDs:
9140 (Human); 67526 (Mouse); 361321 (Rat)
Apg12 (autophagy 12, S. cerevisiae)-like; APG12 autophagy 12-like (S. cerevisiae); APG12; APG12HAPG12; APG12-like; ATG12 autophagy related 12 homolog (S. cerevisiae); ATG12; Autophagy-related protein 12; FBR93; HAPG12; ubiquitin-like protein ATG12; yeast) homolog
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We have 1 review tested in 1 application: Western Blot.