APG8 Antibody - BSA Free

Western Blot: APG8 Antibody [NB600-471] - Western blot using APG8 antibody generated by immunization with recombinant yeast APG8 vs other anti-UBL antibodies against E.coli lysates expressing APG8-GFP fusion protein. Panel A (top) total protein staining using ponceau. Panel B (bottom) specific reaction with APG8 using a 1:4,000 - 1:8,000 dilution of IgG fraction of Rabbit-anti-APG8 (Yeast) followed by 1:15,000 dilution of HRP Goat-a-Rabbit IgG MX. All primaries diluted in TTBS buffer with 5% non-fat milk and incubated with membranes overnight at 4C. E.coli lysate proteins separated by SDS-PAGE using 15% gel. Similar experiments (data not shown) using this antibody showed no reactivity of anti-APG8 with other UBLs, indicating that this antibody 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.

N/A: APG8 Antibody [NB600-471]

• Reactivity: Ye
• Applications: WB, ELISA, IHC, IP
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free

APG8 Antibody - BSA Free Summary

• Description: This antibody, also known as APG8, 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.
  
  Store antibody at 4C prior to restoration. For extended storage aliquot contents and freeze at -20C 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 4C as an undiluted liquid. Dilute only prior to immediate use.
• Immunogen: APG8 antibody was prepared from rabbit serum after repeated immunizations with recombinant yeast APG8 protein.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: ATG8
• Purity: Multi-step

Applications/Dilutions

• Dilutions:
  • ELISA 1:20000-1:100000
  • Immunohistochemistry 1:10 - 1:500
  • Immunohistochemistry-Paraffin 1:10 - 1:500
  • Immunoprecipitation 1:10 - 1:500
  • Western Blot 1:4000-1:8000
• Application Notes: This product 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.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
• Preservative: 0.01% Sodium Azide
• Purity: Multi-step

Alternate Names for APG8 Antibody - BSA Free

• APG8
• Atg8p
• AUT7
• CVT5

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. 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

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: ATG8
• Uniprot:
  • P38182(Yeast)