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ATG9A Overexpression Lysate

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Western Blot: ATG9A Overexpression Lysate (Adult Normal) [NBP2-09258] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for ATG9A.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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ATG9A Overexpression Lysate Summary

Description

ATG9A Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC222568

Accession#: NM_001077198

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
ATG9A

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 (cat# 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
94.3 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 ATG9A Overexpression Lysate

  • APG9 autophagy 9-like 1 (S. cerevisiae)
  • APG9 autophagy 9-like 1
  • APG9L1APG9-like 1
  • ATG9 autophagy related 9 homolog A (S. cerevisiae)
  • autophagy 9-like 1 protein
  • autophagy-related protein 9A
  • FLJ22169
  • mATG9
  • MGD3208

Background

Survival of environmental stress conditions requires the maintenance of cellular homeostasis. To preserve this balance, cells utilize a degradative mechanism known as autophagy. During this process, in response to starvation or other stresses, bulk cytoplasm is non-specifically sequestered within double-membrane vesicles and delivered to the lysosome/vacuole for subsequent degradation and recycling. Atg9 is the only integral membrane protein required for autophagosome formation and is considered a membrane carrier in autophagy-related pathways.

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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Product General Protocols

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Research Areas for ATG9A Lysate (NBP2-09258)

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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.

Mechanisms of Neurodegeneration: Protein aggregation and failure of autophagy
By Michalina Hanzel, PhDIn a series of three blog posts I will briefly explore the major cellular mechanisms responsible for many neurodegenerative disorders. The first, and perhaps the most apparent, is the accumulat...  Read full blog post.

Animal Models to Study Autophagy
By Christina Towers, PhD What is autophagy?Autophagy is the catabolic process that degrades cytoplasmic material via the lysosome. The process of macroautophagy was originally characterized in yeast, where the...  Read full blog post.

Atg9b - a marker for autophagosome induction and assembly
Atg9 is the only essential transmembrane protein involved in cellular autophagy. Autophagy regulates cellular homeostasis by allowing the turnover and recycling of misfolded proteins and damaged organelles. Formation of the double-membrane isolatio...  Read full blog post.

ATG9A - early marker autophagosome assembly
ATG9A is the only essential integral membrane protein involved in autophagy. ATG9A contains six transmembrane domains and initiates the assembly of autophagosomes. The autophagosome is a double-membrane structure that engulfs and eventually degrade...  Read full blog post.

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Bioinformatics

Gene Symbol ATG9A