Autophagy Related 12 (ATG12) antibody

Details for Product No. ABIN116953
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Antigen
Synonyms APG12, APG12L, FBR93, HAPG12, 4931423H11Rik, A330058M13Rik, Apg12l, Atg12l
Reactivity
Yeast (Saccharomyces cerevisiae), Yeast
(105), (43), (40), (12), (12), (12), (4), (1)
Host
Rabbit
(100), (15), (4), (1)
Clonality
Polyclonal
Conjugate
Un-conjugated
(5), (5), (5), (4), (4), (4), (1), (1), (1), (1), (1), (1), (1), (1)
Application
Enzyme Immunoassay (EIA), Western Blotting (WB)
(97), (58), (55), (16), (15), (14), (10), (5), (4), (3), (3), (1), (1)
Pubmed 8 references available
Quantity 0.5 mg
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Catalog No. ABIN116953
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Immunogen This purified antibody was prepared from rabbit serum after repeated immunizations with recombinant yeast APG12 protein.
Specificity Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Rabbit Serum.
Cross-Reactivity (Details) Species reactivity (tested):S. cerevisiae, Yeast
Purification A multi-step process which includes Delipidation, Salt Fractionation and Ion exchange chromatography followed by extensive dialysis against the buffer stated below.
Alternative Name ATG12 / APG12
Background Ubiquitin-like proteins 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. In yeast, autophagy, the delivery of cytoplasmic components to the lysosome/vacuole for degradation, requires a ubiquitin-like protein conjugation system, in which Apg12 is covalently bound to Apg12-Apg5 and Apg16.Synonyms: APG12-like, APG12L, Autophagy-related protein 12, Ubiquitin-like protein ATG12
Gene ID 852518
NCBI Accession NP_009776
UniProt P38316
Application Notes This purified polyclonal antibody reacts with yeast APG12 by Western blot and ELISA. This antibody using the specified conditions may recognize other prominent intrinsicbands (UBLs or conjugates).
Other intrinsic bands are readily detectable at lower dilutions. A 22.1 kDa band corresponding to yeast APG12 is detected. Most yeast cell lysates can be used as a positive control without induction or stimulation. Recommended Dilutions: ELISA: 1/1,000-1/5,000. Western Blot: 1/200. Although not tested, this antibody is likely functional in Immunohistochemistry andImmunoprecipitation.
Other applications not tested.
Optimal dilutions are dependent on conditions and should be determined by the user.
Restrictions For Research Use only
Reconstitution Restore with 0.1 mL of deionized water or equivalent.
Concentration 5.0 mg/mL (by UV absorbance at 280 nm)
Buffer 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2 with 0.01 % (w/v) Sodium Azide as preservative
Preservative Sodium azide
Precaution of Use This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice Avoid repeated freezing and thawing. Dilute only prior to immediate use
Storage 4 °C/-20 °C
Storage Comment Store vial at 2-8 °C prior to restoration. For extended storage add glycerol to 50% and then aliquot contents and freeze at -20 °C or below. Centrifuge product if not completely clear after standing at room temperature. This antibody is stable for one month at 2-8 °C as an undiluted liquid.
Supplier Images
anti-Autophagy Related 12 (ATG12) antibody Immunoblot of APG12 fusion protein. Anti-APG12 antibody generated by immunization with recombinant yeast APG12 was tested by immunoblot against yeast lysates expressing the APG12-GFP fusion protein and other UBL fusion proteins. 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 positions of free GFP or GFP containing recombinant proteins present in each lysate preparation after reaction with a 1:1,000 dilution of ROCKLAND's anti-GFP (code # 600-101-215) followed by reaction with a 1:15,000 dilution of HRP Donkey-a-Goat IgG MX (code # 605-703-125). Panel C shows specific reaction with APG12 using a 1:2,000 dilution of ROCKLAND's IgG fraction of Rabbit-anti-APG12 (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. Yeast lysate proteins were separated by SDS-PAGE using a 15% gel. This data indicates that anti-APG12 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.
anti-Autophagy Related 12 (ATG12) antibody (2) Conjugation pathways for ubiquitin and ubiquitin-like modifiers (UBLs). Most modifiers mature by proteolytic processing from inactive precursors (a, amino acid). Arrowheads point to the cleavage sites. Ubiquitin is expressed either as polyubiquitin or as a fusion with ribosomal proteins. Conjugation requires activating (E1) and conjugating (E2) enzymes that form thiolesters (S) with the modifiers. Modification of cullins by RUB involves SCF(SKP1/cullin-1/F-box protein) /CBC(cullin- 2/elongin B/elonginC) -like E3 enzymes that are also involved in ubiquitination. In contrast to ubiquitin, the UBLs do not seem to form multi-UBL chains. UCRP(ISG15) resembles two ubiquitin moieties linked head-to-tail. Whether HUB1 functions as a modifier is currently unclear. APG12 and URM1 are distinct from the other modifiers because they are unrelated in sequence to ubiquitin. Data contributed by S.Jentsch, see references below.
Background publications Krysko, Samoylenko, Polishchuk et al.: "RGD mimetics containing phthalimidine fragment as novel ligands of fibrinogen receptor." in: Bioorganic & medicinal chemistry letters, Vol. 21, Issue 19, pp. 5971-4, 2011 (PubMed).

Cohn, Goddijn, Middeldorp et al.: "Recurrent miscarriage and antiphospholipid antibodies: prognosis of subsequent pregnancy." in: Journal of thrombosis and haemostasis : JTH, Vol. 8, Issue 10, pp. 2208-13, 2010 (PubMed).

Mizushima, Kuma, Kobayashi et al.: "Mouse Apg16L, a novel WD-repeat protein, targets to the autophagic isolation membrane with the Apg12-Apg5 conjugate." in: Journal of cell science, Vol. 116, Issue Pt 9, pp. 1679-88, 2003 (PubMed).

Kuma, Mizushima, Ishihara et al.: "Formation of the approximately 350-kDa Apg12-Apg5.Apg16 multimeric complex, mediated by Apg16 oligomerization, is essential for autophagy in yeast." in: The Journal of biological chemistry, Vol. 277, Issue 21, pp. 18619-25, 2002 (PubMed).

Suzuki, Kirisako, Kamada et al.: "The pre-autophagosomal structure organized by concerted functions of APG genes is essential for autophagosome formation." in: The EMBO journal, Vol. 20, Issue 21, pp. 5971-81, 2001 (PubMed).

Jentsch, Pyrowolakis: "Ubiquitin and its kin: how close are the family ties?" in: Trends in cell biology, Vol. 10, Issue 8, pp. 335-42, 2000 (PubMed).

Mizushima, Sugita, Yoshimori et al.: "A new protein conjugation system in human. The counterpart of the yeast Apg12p conjugation system essential for autophagy." in: The Journal of biological chemistry, Vol. 273, Issue 51, pp. 33889-92, 1999 (PubMed).

Liakopoulos, Doenges, Matuschewski et al.: "A novel protein modification pathway related to the ubiquitin system." in: The EMBO journal, Vol. 17, Issue 8, pp. 2208-14, 1998 (PubMed).

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