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ACOT8 Overexpression Lysate

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Western Blot: ACOT8 Overexpression Lysate (Adult Normal) [NBP2-07748] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for ACOT8.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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ACOT8 Overexpression Lysate Summary

Description

ACOT8 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC214175

Accession#: NM_005469

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
ACOT8

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
35.7 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 ACOT8 Overexpression Lysate

  • ACTEIII
  • acyl-CoA thioesterase 8HNAACTE
  • Choloyl-coenzyme A thioesterase
  • EC 3.1.2.27
  • hACTEIII
  • HIV-Nef-associated acyl-CoA thioesterase
  • hTEcholoyl-CoA hydrolase
  • long-chain fatty-acyl-CoA hydrolase
  • palmitoyl-CoA hydrolase
  • peroxisomal acyl-CoA thioesterase 1
  • peroxisomal acyl-CoA thioesterase
  • Peroxisomal acyl-coenzyme A thioester hydrolase 1
  • Peroxisomal long-chain acyl-CoA thioesterase 1
  • PTE-1
  • PTE1HIV-Nef associated acyl-CoA thioesterase
  • PTE2
  • PTE-2hACTE-IIIacyl-coenzyme A thioesterase 8
  • Thioesterase II
  • thioesterase III

Background

ACOT8 is encoded by this gene is a peroxisomal thioesterase that appears to be involved more in the oxidation of fatty acids rather than in their formation. The encoded protein can bind to the human immunodeficiency virus-1 protein Nef, and mediate Nef-induced down-regulation of CD4 in T-cells. Multiple transcript variants encoding several different isoforms have been found for this gene. [provided by RefSeq]

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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Bioinformatics

Gene Symbol ACOT8