Human/Mouse ATG12 Antibody

(1 Review)
  
  • Species Reactivity
    Human, Mouse
  • Specificity
    Detects human ATG12 in direct ELISAs, and human and mouse ATG12 in Western blots.
  • Source
    Monoclonal Mouse IgG2B Clone # 628902
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant human ATG12
    Asp48-Ala187
    Accession # O94817
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.1 µg/mL
    See below
  • Immunocytochemistry
    8-25 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
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.
Immunocytochemistry
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
  • Reconstitution
    Sterile PBS to a final concentration of 0.5 mg/mL.
  • Shipping
    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.
Background: ATG12
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.
  • Long Name:
    ATG12 Autophagy Related 12 Homolog
  • Entrez Gene IDs:
    9140 (Human); 67526 (Mouse); 361321 (Rat)
  • Alternate Names:
    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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Average Rating: 5 (Based on 1 review)

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We have 1 review tested in 1 species: Human.
We have 1 review tested in 1 application: Western Blot.

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 WB Human Anonymous 11/16/2017
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Summary

ApplicationWestern Blot
Sample TestedCell Lysates
SpeciesHuman

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