Lipase A Overexpression Lysate

Western Blot: Lipase A Overexpression Lysate (Adult Normal) [NBL1-12546] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Lipase A.

• Reactivity: Hu
• Applications: WB

Lipase A Overexpression Lysate Summary

• Description:

  Lipase A Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC201637
  
  Accession#: NM_000235
  
  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: LIPA

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: 43 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 Lipase A Overexpression Lysate

• Acid cholesteryl ester hydrolase
• CESD
• Cholesteryl esterase
• EC 3.1.1
• EC 3.1.1.13
• LALCESDcholesterol ester hydrolase
• LIPA
• Lipase A
• lipase A, lysosomal acid, cholesterol esterase
• lysosomal acid lipase
• lysosomal acid lipase/cholesteryl ester hydrolase
• Sterol esterase

Background

Lipase A, also known as LIPA or Lysosomal acid lipase/cholesteryl ester hydrolase, has a 399 amino acid long isoform that is 45 kDa and a short 343 amino acid isoform that is 39 kDa; lysosome located; is very important for the intracellular hydrolysis of cholesteryl esters and triglycerides that have been internalized via receptor-mediated endocytosis of lipoprotein particles, and also in mediating the effect of LDL (low density lipoprotein) uptake on suppression of hydroxymethylglutaryl-CoA reductase and activation of endogenous cellular cholesteryl ester formation. This protein is currently being studied for involvement in the following diseases and disorders: cholesteryl ester storage disease, wolman disease, cholesterol, cholesterol ester storage disease, lysosomal storage disease, coronary heart disease, hepatitis c, splenomegaly, hepatitis b, hepatitis, hyperlipidemia, hypercholesterolemia, tuberculosis, obesity atherosclerosis, alzheimer's disease, neisseria meningitides, lipid storage disease, lipodystrophy, pandas, wolman disease, and blepharitis. The LIPA protein has been shown to interact with HIST1H4A, HIST1H4B, HIST1H4C, SOAT1, and HIST1H4E in cholesterol and sphingolipids transport / distribution to the intracellular membrane compartments (normal and CF), steroid biosynthesis, and lysosome 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.

Bioinformatics

• Gene Symbol: LIPA