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Primary Antibodies  >  Ubiquitin and UBL Antibodies

APG8 Antibody

Rabbit Polyclonal
NCI Collaboration
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Western blot of APG8 fusion protein.  Anti-APG8 antibody generated by immunization with recombinant yeast APG8 was tested by western blot with other anti-UBL antibodies against E.coli lysates expressing the APG8-GFP fusion protein.  All UBLs possess limited homology to Ubiquitin and to each other, therefore it is important to know the degree of reactivity of each antibody against each UBL.   Panel A shows total protein staining using ponceau.  Panel B shows specific reaction with APG8 using a 1:4,000 and 1:8,000 dilution of Rockland's IgG fraction of Rabbit-anti-APG8 (Yeast) followed by reaction with a 1:15,000 dilution of HRP Goat-a-Rabbit IgG MX (code # 611-103-122).  All primary antibodies were diluted in TTBS buffer supplemented with 5% non-fat milk and incubated with the membranes overnight at 4° C.  E.coli lysate proteins were separated by SDS-PAGE using a 15% gel.  Similar experiments (data not shown), where other UBL fusion proteins were separated and probed with this antibody showed no reactivity of anti-APG8 with other UBLs.  This data indicates that anti-APG8 is highly specific and does not cross react with other UBLs.   A chemiluminescence system was used for signal detection (Roche).  Other detection systems will yield similar results.  Data contributed by M. Malakhov, www.lifesensors.com, personal communication.
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500 µg
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APG8 Antibody Properties

Anti-APG8 (RABBIT) Antibody - 200-401-439
Target Species
Known Cross Reactivity
ELISA : 1:20,000 - 1:100,000
Western Blot : 1:4,000 - 1:8,000
Other Dilution: User Optimized
Physical State
Shipping Condition
5.0 mg/mL by UV absorbance at 280 nm
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Reconstitution Volume
100 µL
Reconstitution Buffer
Restore with deionized water (or equivalent)
0.01% (w/v) Sodium Azide
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APG8 Antibody Description

Anti-ATG8 antibody is an Ubiquitin-like protein (UBL) antibody. UBL proteins and antibodies fall into two classes: the first class, ubiquitin-like modifiers (UBLs) function as modifiers in a manner analogous to that of ubiquitin.  Examples of UBLs are SUMO, Rub1 (also called Nedd8), Apg8 and Apg12.  Proteins of the second class include parkin, RAD23 and DSK2, are designated ubiquitin-domain proteins (UDPs).  These proteins contain domains that are related to ubiquitin but are otherwise unrelated to each other.  In contrast to UBLs, UDPs are not conjugated to other proteins.  Apg8 is required for autophagy (intracellular bulk protein degradation) in yeast.  Starved yeast cells take up their own cytoplasm into vacuoles through autophagic bodies.   Autophagic bodies form a double-membraned structure called the autophagosome, which subsequently fuses with the vacuole/lysosome. This process similar in mammals.  Two sets of genes, APG and AUT, have been identified with this process, and are responsible for two ubiquitin-like systems Apg12 and Apg8, respectively.  Apg12 is synthesized in its mature form and seems to have one target, Apg5.  Almost all Apg12 molecules are conjugated with Apg5.  Aut2/Apg4 processes the Apg8/Aut7 system at its carboxy-terminal region.  Apg8 exists in two forms, one is membrane bound through a phospholipid. Lipidation/ activation of Apg8 is mediated by Apg7 and transferred to Apg3 and finally forms a conjugate with phosphatidyl-ethanolamine (PE). Apg4 cleaves Apg8–PE, releasing Apg8 from membrane.  Morphological studies show that Apg8 localizes on the membrane of intermediate structures of the autophagosome; this transient association seems to be essential for formation of the autophagosome. Both Apg12 and Apg8 are highly conserved, with apparent homologues in the worm, mammals and plants. In higher eukaryotes, Apg8 consists of a multigene family. Useful in Cell Biology, Microbiology and Signal Transduction research.
ATG8 Antibody, Autophagy-related protein 8, Autophagy-related ubiquitin-like modifier ATG8, Cytoplasm to vacuole targeting protein 5
This purified antibody was prepared from rabbit serum after repeated immunizations with recombinant yeast APG8 protein.
Immunogen Type
Recombinant Protein
Storage Condition
Store antibody at 4° C prior to restoration.   For extended storage aliquot contents and freeze at -20° C or below.  Avoid cycles of freezing and thawing.  Centrifuge product if not completely clear after standing at room temperature.  This product is stable for several weeks at 4° C as an undiluted liquid.  Dilute only prior to immediate use.
Application Note
This purified polyclonal antibody reacts with yeast APG8 by western blot and ELISA. Although not tested, this antibody is likely functional in immunohistochemistry and immunoprecipitation.   This antibody using the specified conditions may recognize other prominent intrinsic bands (UBLs or their conjugates).  Other intrinsic bands are readily detectable in yeast lysates at lower antibody dilutions. For immunoblotting a 14 kDa band corresponding to yeast APG8 is detected.  Most yeast cell lysates can be used as a positive control without induction or stimulation. 
Anti-ATG8 antibody is also known as Anti-APG8 and is an IgG fraction antibody purified from monospecific antiserum by a multi-step process which includes delipidation, salt fractionation and ion exchange chromatography followed by extensive dialysis against the buffer stated above.  Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Rabbit Serum.
Disclaimer Note-General
This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 5199, Limerick, Pennsylvania, USA.
General Reference
Kirisako T, Baba M, Ishihara N, Miyazawa K, Ohsumi M, Yoshimori T, Noda T, Ohsumi Y. (1999) Formation process of autophagosome is traced with Apg8/Aut7p in yeast. J Cell Biol.;147(2):435-46. Ohsumi, Y. (2001) Molecular dissection of autophagy: two ubiquitin-like systems. Nature Reviews Molecular Cell Biology 2, 211 -216. Liakopoulos D et al. (1998). A novel protein modification pathway related to the ubiquitin system. EMBO J. 15;17(8):2208-14.
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