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ATG16L1 Overexpression Lysate

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Western Blot: ATG16L1 Overexpression Lysate (Adult Normal) [NBP2-06180] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for ATG16L1.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

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ATG16L1 Overexpression Lysate Summary

Description

ATG16L1 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC216728

Accession#: NM_030803

Protein Tag: C-MYC/DDK

You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.
Gene
ATG16L1

Applications/Dilutions

Dilutions
  • Western Blot
Application Notes
This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.

Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
Theoretical MW
68.1 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

Storage
Store at -80C. Avoid freeze-thaw cycles.
Buffer
RIPA buffer

Lysate Details for Array

Type
Overexpression

Notes

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

Alternate Names for ATG16L1 Overexpression Lysate

  • APG16 autophagy 16-like (S. cerevisiae)
  • APG16L beta
  • APG16LFLJ10828
  • APG16-like 1
  • ATG16 autophagy related 16-like (S. cerevisiae)
  • ATG16 autophagy related 16-like 1 (S. cerevisiae)
  • ATG16A
  • ATG16L
  • autophagy-related protein 16-1
  • FLJ00045
  • FLJ10035
  • FLJ22677
  • IBD10
  • WD repeat domain 30
  • WDR30

Background

Macroautophagy is the major inducible pathway for the general turnover of cytoplasmic constituents in eukaryotic cells, it is also responsible for the degradation of active cytoplasmic enzymes and organelles during nutrient starvation. Macroautophagy involves the formation of double-membrane bound autophagosomes which enclose the cytoplasmic constituent targeted for degradation in a membrane bound structure, which then fuse with the lysosome (or vacuole) releasing a single-membrane bound autophagic bodies which are then degraded within the lysosome (or vacuole). The APG12-APG5-APG16L complex is essential for the elongation of autophagic isolation membranes. This complex initially associates in uniform distribution with small vesicle membranes. During membrane elongation, the complex partitions, with a great concentration building on the outer side of the isolation membrane. Upon completion of the formation of the autophagosome, the APG12-APG5-APG16L dissociates from the membrane. There are 5 isoforms of this protein (in human), with theoretical molecular weights ranging from ~19-68 kDa.

Mutations in the ATG16L1 proitein have been associated with Crohn's disease.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

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Liver ASK1 activates autophagy to protect against hepatic fat accumulation, non-alcoholic steatohepatitis and fibrosis
By Jamshed Arslan, Pharm. D., PhD. The most common chronic liver disorder worldwide is non-alcoholic fatty liver disease (NAFLD). This obesity-linked disorder can manifest as hepatic fat accumulation (steatosis) wit...  Read full blog post.

ATG16L1 - a key player in the development of the autophagosome
Like apoptosis, autophagy is a highly regulated physiologic process that involves cellular degradation and recycling of organelles and macromolecules. Autophagy is a survival mechanism induced by states of stress, starvation, and infection. A do...  Read full blog post.

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Bioinformatics

Gene Symbol ATG16L1