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).
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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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