General description
We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry. This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency".
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ZooMAb® antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb® antibodies are reliably available and ready to ship when you need them.
Specificity
Clone 1I5 is a ZooMAb® rabbit recombinant monoclonal antibody that specifically detects Autophagy-related protein 16-1 (ATG16L). It targets an epitope within 25 amino acids from the N-terminal half. It detects both isoforms.
Immunogen
KLH-conjugated linear peptide corresponding to 25 amino acids from the N-terminal half of human Autophagy-related protein 16-1 (ATG16L).
Application
Quality Control Testing
Evaluated by Western Blotting in PANC-1 cell lysate.
Western Blotting Analysis: A 1:1,000 dilution of this antibody detected ATG16L in PANC-1 cell lysate.
Tested Applications
Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected ATG16L in Rat testis tissue lysate.
Affinity Binding Assay: A representative lot of this antibody bound ATG16L peptide with a KD of 2.4 x 10-6 in an affinity binding assay.
Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected ATG16L in HeLa cells.
Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected ATG16L in human testis tissue sections.
Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.
Target description
Autophagy-related protein 16-1 (UniProt: Q676U5; also known as APG16-like 1) is encoded by the ATG16L1 (also known as APG16L, UNQ9393/PRO34307) gene (Gene ID: 55054) in human. Autophagy is a highly regulated process that is involved in cell growth, development, and cell death. In this process cytoplasmic components are delivered to the lysosome in a very systematic manner for degradation. Dysregulated autophagy has been reported in several human pathophysiologic conditions. ATG16L1 is a homodimeric peripheral protein that plays an essential role in both canonical and non-canonical autophagy. It is responsible for the correct targeting of LC3 to form autophagosome membranes. It contains an N‐terminal domain that associates with ATG5 and ATG12, a central FIP200 and WIPI2b binding domain, and a C‐terminal WD40 domain (aa 320-607) with seven WD repeats. The ATG16L1-ATG5‐ATG12 complex has an E3‐Ubiquitin ligase‐like enzymatic activity, which is reported to be essential for lipidation of LC3 and for targeting of LC3 to membranes during canonical and non‐canonical autophagy. The central region in ATG16L1 that has binding sites for both FIP200 and WIPI2b is known as the FIP200 binding domain. Deletion of this domain is shown to prevent ATG16L1 recruitment to form autophagosomes and inhibit the canonical autophagy response to amino acid starvation and bacterial infection. C-terminal WD repeats containing domain is essential for LC3 recruitment to endo-lysosomal membranes during non-canonical autophagy but is shown to be dispensable for canonical autophagy. ATG16L1 is also reported to regulate mitochondrial antiviral signaling (MAVS)-dependent type I interferon production and negatively regulates NOD1- and NOD2-driven inflammatory cytokine response. Five isoforms of ATG16L1 have been described that are produced by alternative splicing. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Fletcher, K., et al. (2018). EMBO J. 37(4); e97840; Nishimura, T., et al. (2013). EMBO Rep. 14(3); 284-291; Fujita, N., et al. (2008). Mol. Biol. Cell. 19(5); 2092-2100).
Physical form
Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 µL.
Reconstitution
300 µg/mL after reconstitution at 25 µL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.
Storage and Stability
Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 µL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws.
Other Notes
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Legal Information
ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.