Detection of Human and Mouse GABARAP by Western Blot.
Western blot shows lysates of A172 human glioblastoma cell line and NIH‑3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 50 µM Chloroquine for 20 hours. PVDF membrane was probed with 2 µg/mL of Rat Anti-Human GABARAP Monoclonal Antibody (Catalog # MAB8574) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). Specific bands were detected for GABARAP at approximately 14 & 16 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 7.
Preparation and Storage
Reconstitute at 0.5 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
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.
gamma -Aminobutyric Acid Receptor-associated Protein (GABARAP), also known as Apg8p1, is 117 amino acid (aa) protein with a predicted molecular weight of 14 kDa. It is a member of the GABARAP subfamily of the Autophagy-related 8 (Atg8) family of proteins (1). The GABARAP subfamily also includes GABARAPL1. GABARAP/Apg8p1 has 100% aa sequence identity with its mouse and rat orthologs and is orthologous to yeast Atg8. Atg8 family members show structural similarity with Ubiquitin, but lack aa sequence similarity. GABARAP/Apg8p1 was first described for its putative involvement GABA A Receptor trafficking (2,3). However, it is best known for its role in autophagy (1,4). GABARAP/Apg8p1 covalently attaches to phosphatidylethanolamine (PE) in the phagophore (autophagosome precursor) membrane using a Ubiquitin-like conjugation system that includes Ubiquitin-activating (E1)-, Ubiquitin-conjugating (E2)-, and Ubiquitin Ligase (E3)-like enzymes. Here it is involved in the later stages of autophagosome maturation (4,5). It may also be involved in cargo recruitment to autophagosomes (1).
Shpilka, T. et al. (2011) Genome Biol. 12:226.
Wang, H. et al. (1999) Nature 397:69.
Leil, T.A. et al. (2004) J. Neurosci. 24:11429.
Weidberg, H. et al. (2010) EMBO J. 29:1792.
Weidberg, H. et al. (2011) Ann. Rev. Biochem. 80:125.
gamma-Aminobutyric Acid Type A Receptor Associated Protein
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