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ATG4B Overexpression Lysate

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Western Blot: ATG4B Overexpression Lysate (Adult Normal) [NBP2-09850] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for ATG4B.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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ATG4B Overexpression Lysate Summary

Description

ATG4B Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC200289

Accession#: NM_013325

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
ATG4B

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
44.1 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 ATG4B Overexpression Lysate

  • APG4 autophagy 4 homolog B (S. cerevisiae)
  • APG4 autophagy 4 homolog B
  • Apg4B
  • ATG4 autophagy related 4 homolog B (S. cerevisiae)
  • ATG4B
  • AUTL1
  • AUTL1MGC1353
  • AUT-like 1 cysteine endopeptidase
  • autophagin-1
  • Autophagy-related cysteine endopeptidase 1
  • Autophagy-related protein 4 homolog B
  • cysteine protease ATG4B
  • DKFZp586D1822
  • EC 3.4.22
  • EC 3.4.22.-
  • hAPG4B
  • KIAA0943

Background

FUNCTION: Cysteine protease required for autophagy, which cleaves the C-terminal part of either MAP1LC3, GABARAPL2 or GABARAP, allowing the liberation of form I. A subpopulation of form I is subsequently converted to a smaller form (form II). Form II, with a revealed C-terminal glycine, is considered to be the phosphatidylethanolamine (PE)-conjugated form, and has the capacity for the binding to autophagosomes. ENZYME REGULATION: Inhibited by N-ethylmaleimide. SUBCELLULAR LOCATION: Cytoplasm (Probable). ALTERNATIVE PRODUCTS: 5 named isoforms produced by alternative splicing.

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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Blogs on ATG4B.

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.


  Read full blog post.

ATG4B - a cysteine protease involved in autophagosome elongation
Autophagy can be broken down into 4 main stages: phagophore nucleation, autophagosome elongation, autophagosome docking and fusion with a lysosome, and vesicle breakdown and degradation. ATG4B is one of four ATG4 homologs (ATG4A, ATG4B, ATG4C, and ...  Read full blog post.

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Bioinformatics

Gene Symbol ATG4B