ATG8 Antibody

SKU: orb344618

Description

APG8 antibody

Images & Validation

• Tested Applications: ELISA, WB
• Dilution Range: ELISA: 1:20,000 - 1:100,000, WB: 1:4,000 - 1:8,000
• Reactivity: Yeast
• Application Notes:
  Anti-Apg8 purified polyclonal antibody reacts with yeast APG8 by western blot and ELISA. 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. Although not tested, this antibody is likely functional in immunohistochemistry and immunoprecipitation.

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Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Immunogen: Rabbit Anti-Apg8 antibody was prepared from rabbit serum after repeated immunizations with recombinant yeast Apg8 protein.
• Target: ATG8
• Purity: Anti-Apg8 antibody, also known as Atg8, 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.
• Conjugation: Unconjugated

Storage & Handling

• Storage: 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.
• Form/Appearance: Lyophilized
• Buffer/Preservatives: Preservative: 0.01% (w/v) Sodium Azide. Stabilizer: None; Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
• Concentration: 5.0 mg/mL
• Expiration Date: 12 months from date of receipt.
• Disclaimer: For research use only

Alternative Names

rabbit anti-APG8 Antibody, APG-8, APG 8, ATG-8, ATG 8, Rabbit Anti-ATG8 Antibody, Autophagy-related protein 8, Autophagy-related ubiquitin-like modifier ATG8, Cytoplasm to vacuole targeting protein 5, AUT7, CVT5, YBL078C, YBL0732

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.

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 (top) shows total protein staining using ponceau. Panel B (bottom) shows specific reaction with APG8 using a 1:4000 and 1:8000 dilution of Biorbyt's IgG fraction of Rabbit-anti-APG8 (Yeast) followed by reaction with a 1:15000 dilution of HRP Goat-a-Rabbit IgG MX (orb347654). 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.

Quick Database Links

Gene Symbol

ATG8

UniProt

P38182

RefSeq:6319393