Product DescriptionMouse anti-Autophagy-related protein 5 (ATG5) Monoclonal Antibody (Unconjugated), suitable for WB, ELISA.
Application(s)ELISA, WB
Application DetailsThis antibody is recommended for WB, sandwich ELISA, Flow Cytometry (~2 ug per 10^6 cells). Biosensis recommends optimal dilutions/concentrations should be determined by the end user.
TargetAutophagy-related protein 5 (ATG5)
SpecificitySpecificity has been confirmed by WB and direct ELISA against the antigen. Human. Other species have not been tested.
Target Host SpeciesHuman
Species ReactivityHuman
Antibody HostMouse
Antibody TypeMonoclonal
Antibody IsotypeIgG2a, kappa
Clone Name4B2
ConjugateUnconjugated
Immunogen DescriptionPartial recombinant human ATG5 (176-275) with a GST tag.
Purity DescriptionProtein G purified immunoglobulin
FormatLyophilized from PBS pH 7.2
Reconstitution InstructionsSpin vial briefly before opening. Reconstitute in 100 µL sterile-filtered, ultrapure water. Centrifuge to remove any insoluble material.
Storage InstructionsAfter reconstitution keep aliquots at -20°C for higher stability or at 2-8°C with an appropriate antibacterial agent. Glycerol (1:1) may be added for additional stability. Avoid repetitive freeze/thaw cycles.
Batch NumberPlease see item label.
Expiration Date12 months after date of receipt (unopened vial).
Alternative NamesAutophagy protein 5; APG5-like; Apoptosis -specific protein; ATG5; APG5L; ASP; APG6
Autophagy is a process of bulk protein degradation in which cytoplasmic components, including organelles, are enclosed in double-membrane structures called autophagosomes and delivered to lysosomes or vacuoles for degradation. ATG5 conjugates to ATG12 and associates with isolation membrane to form cup-shaped isolation membrane and autophagosome. The conjugate detaches from the membrane immediately before or after autophagosome formation is completed. ATG5 may also play an important role in the apoptotic process, possibly within the modified cytoskeleton. Its expression is a relatively late event in the apoptotic process, occurring downstream of caspase activity. It localise to the cytoplasm and it co-localise with nonmuscle actin. ATG5 is ubiquitously expressed. Its expression is dramatically increased in apoptotic cells. Conjugation to ATG12 is essential for autophage but it is not required for association with isolation membrane.
A: Specific staining of ATG5 expressed in human neuroblastoma SH-SY5Y cell line by Flow Cytometry using cat # M-882-100. Fixing and Permeabilization of cells: Absolute methanol (10 minutes in ice) and 0.1% Tween-20 in PBS, Blocking: 200 µg/mL Normal Sheep IgG (20 minutes), Primary antibody: Mouse Monoclonal antibody to ATG5 (2 μg per ~10^6 cells) for 30 minutes at room temperature, Secondary antibody: Goat anti-mouse PE labeled secondary antibody (1:100 dilution), incubation for 20 minutes in dark at room temperature. Negative control: Non-specific Control IgG, clone X63 (cat # M-1249-200, black dashed). Data and results were generated using Orflo MoxiflowTM instrument and protocols. B: Western blot detection of GST tagged recombinant human ATG5. Note the GST tag alone is 26 kDa. C: Sandwich ELISA using anti-ATG5 monoclonal antibody as a capture antibody. The detection limit for GST tagged recombinant ATG5 protein is 0.1 ng/ml.
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