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Primary Antibodies  >  Ubiquitin and UBL Antibodies
 

APG12 Antibody

Rabbit Polyclonal
Code

200-401-437
Size

500 µg
Price

$309.00
Availability

Ships next business day.
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Western blot of APG12 fusion protein.  Anti-APG12 antibody generated by immunization with recombinant yeast APG12 was tested by western blot 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, Lifesensors Inc., personal communication.
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Synonyms: APG12-like antibody, APG12L antibody, ATG12 antibody, ATG12 autophagy related 12 homolog (S. cerevisiae) antibody, ATG12 autophagy related 12 homolog antibody, Autophagy 12 antibody, Autophagy-related protein 12 antibody, HAPG12 antibody

APG12 Antibody Properties

Description
Anti-APG12 (RABBIT) Antibody - 200-401-437
Host
Rabbit
Target Species
Yeast
Known Cross Reactivity
yeast
Clonality
Polyclonal
Application
ELISA : 1:1,000 - 1:5,000
Western Blot : 1:500 - 1:2,000
Other Dilution: User Optimized
Physical State
Lyophilized
Shipping condition
Ambient
Format
IgG
Label
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
Reconstitution Volume
100 µL
Reconstitution Buffer
Restore with deionized water (or equivalent)
Preservative
0.01% (w/v) Sodium Azide
Stabilizer
None
GeneName
ATG12
Download CofA
Lot#  

APG12 Antibody Description

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.
Immunogen
This purified antibody was prepared from rabbit serum after repeated immunizations with recombinant yeast APG12 protein.
Immunogen Type
Recombinant Protein
Storage Condition
Store vial 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 yeast APG12 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.
Purity/Specifity
This product 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 326, Gilbertsville, Pennsylvania, USA.
General Reference
Kuma A, Mizushima N, Ishihara N, Ohsumi Y. (2002) Formation of the approximately 350-kDa Apg12-Apg5.Apg16 multimeric complex, mediated by Apg16 oligomerization, is essential for autophagy in yeast. J Biol Chem. 24;277(21):18619-25. N.Mizushima et al. (2003) Mouse Apg16L, a novel WD-repeat protein, targets to the autophagic isolation membrane with the Apg12-Apg5 conjugate. J.Cell Science 116, 1679-1688. Suzuki K, Kirisako T, Kamada Y, Mizushima N, Noda T, Ohsumi . (2001) The pre-autophagosomal structure organized by concerted functions of APG genes is essential for autophagosome formation. EMBO J ;20:5971-5981.
Specific Reference
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Product Type
Proteins and Peptides;
Catalog Number
000-001-487

Product Type
Primary Antibodies;
Applications
ELISA, WB, IHC
Reacts With
yeast
Catalog Number
200-301-428

Product Type
Primary Antibodies;
Applications
ELISA, WB
Reacts With
yeast
Catalog Number
200-401-427

Product Type
Primary Antibodies;
Applications
ELISA, WB
Reacts With
yeast
Catalog Number
200-401-428

Product Label

 

Description
Unconjugated
Color
Conjugation Reference
Molecular Weight
Excitation Wavelength
Conjugation Chemistry