Human ATP citrate lyase/ACLY Baculovirus-Insect cells Overexpression Lysate

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Human ATP citrate lyase/ACLY Baculovirus-Insect cells Overexpression Lysate: Product Information

Product Description
This Human ATP citrate lyase/ACLY overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of ATP citrate lyase/ACLY protein (Cat: 11769-H07B) from the overexpression lysate was verified.
Expression Host
Baculovirus-Insect cells
Species
Human
Sequence Information
A DNA sequence encoding the human ACLY (P53396) (Met 1-Met 1101) was expressed, with a polyhistidine tag at the N-terminus.
Molecule Mass
The recombinant human ACLY consists of 1120 amino acids and has a calculated molecular mass of 123 kDa. It migrates as an approximately 110 kDa band in SDS-PAGE under reducing conditions.

Human ATP citrate lyase/ACLY Baculovirus-Insect cells Overexpression Lysate: Usage Guide

Preparation Method
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Lysis Buffer
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Recommend Usage
1.  Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube. 2.  Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.
Sample Buffer
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Stability & Storage
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Human ATP citrate lyase/ACLY Baculovirus-Insect cells Overexpression Lysate: Alternative Names

Human ACL Overexpression Lysate; Human ATPCL Overexpression Lysate; Human CLATP Overexpression Lysate

ATP citrate lyase/ACLY Background Information

ATP citrate lyase, also known as Acly or Acl, is the primary enzyme responsible for the synthesis of cytosolic acetyl-CoA in many tissues. The enzyme is composed of two polymer chains which are polypeptides in human. ATP citrate lyase is responsible for catalyzing the conversion of citrate and CoA into acetyl-CoA and oxaloacetate, along with the hydrolysis of ATP. A definitive role for ATP citrate lyase in tumorigenesis has emerged from ATP citrate lyase RNAi and chemical inhibitor studies, showing that ATP citrate lyase inhibition limits tumor cell proliferation and survival and induces differentiation in vitro. In vivo, it reduces tumor growth leading to a cytostatic effect and induces differentiation.
Full Name
ATP citrate lyase
References
  • Kim W, et al. (2006) Both subunits of ATP-citrate lyase from Chlorobium tepidum contribute to catalytic activity. J Bacteriol. 188 (18) : 6544-52.
  • Ki SW, et al. (2000) Radicicol binds and inhibits mammalian ATP citrate lyase. J Biol Chem. 275 (50) : 39231-6.
  • Schneider K, et al. (2000) Biosynthesis of the prosthetic group of citrate lyase. Biochemistry. 39 (31) : 9438-50.
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