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SOAT1 Overexpression Lysate

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Western Blot: ACAT Overexpression Lysate (Adult Normal) [NBP2-10868] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for ACAT.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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SOAT1 Overexpression Lysate Summary

Description

SOAT1 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC205774

Accession#: NM_003101

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
SOAT1

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
64.6 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 SOAT1 Overexpression Lysate

  • ACACT
  • ACACT1
  • ACAT-1
  • ACATACAT1
  • acyl-Coenzyme A: cholesterol acyltransferase
  • Acyl-coenzyme A:cholesterol acyltransferase 1
  • Cholesterol acyltransferase 1
  • EC 2.3.1.26
  • RP11-215I23.2
  • SOAT1
  • STATSOAT
  • sterol O-acyltransferase (acyl-Coenzyme A: cholesterol acyltransferase) 1
  • sterol O-acyltransferase 1

Background

Acely-CoA:cholestorl acyltransferase (ACAT) is an integral membrane enzyme that belongs to type-B carboxylesterase/lipase family. ACAT catalyzes the formation of cholesteryl esters from cholesterol and long-chain fatty-acyl-coenzyme A (1). ACAT also function as a regulator of intracellular cholesterol homeostasis at single cell level. Two isoform of ACAT exists, ACAT1 and ACAT2, are expressed in liver and intestine. However, ACAT1 is only found in macrophages and SMCs (2). The accumulation of cholesteryl esters produced by ACAT in macrophages can lead to foam cell formation, an indication of early stage atherosclerosis. ACAT activity has been linked to Alzehimer's Diease, condition caused by excess amount of amyloid beta-peptide (Abeta). Atherogenic lipoproteins activate ACAT activity, where active ACAT can modulate the level of Abeta (3).

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 SOAT1