Clusterin/APOJ Overexpression Lysate

Western Blot: APOJ / Clusterin Overexpression Lysate (Adult Normal) [NBP2-04577] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for APOJ / Clusterin.

• Reactivity: Hu
• Applications: WB

Clusterin/APOJ Overexpression Lysate Summary

• Description:

  Clusterin/APOJ Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC203941
  
  Accession#: NM_203339
  
  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: CLU

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: 50 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 Clusterin/APOJ Overexpression Lysate

• 40
• 40, sulfated glycoprotein 2
• Aging-associated gene 4 protein
• aging-associated protein 4
• APOJ
• apo-J
• Apolipoprotein J
• CLI
• CLIclusterin (complement lysis inhibitor, SP-40
• CLU
• Clusterin
• Complement cytolysis inhibitor
• complement lysis inhibitor
• Complement-associated protein SP-40
• Ku70-binding protein 1
• KUB1SGP2
• MGC24903
• NA1/NA2
• SGP-2
• SP-40
• sulfated glycoprotein 2
• Testosterone-repressed prostate message 2
• testosterone-repressed prostate message 2, apolipoprotein J)
• TRPM-2
• TRPM-2TRPM2

Background

Clusterin, also designated complement lysis inhibitor (CLI), apolipoprotein J (APOJ), sulfated glycoprotein 2 (SGP2), SP-40 and testosterone-repressed prostate message 2 (TRPM2), is a secretory, heterodimeric glycoprotein that influences immune regulation, cell adhesion, transformation, lipid transportation, tissue remodeling, membrane recycling and cell-cell interactions. Clusterin is synthesized as a 449 amino acid poly-peptide that is posttranslationally cleaved at an internal bond between Arg 227 and Ser 228. Two subunits, alpha and beta, are associated through disulfide bonds. The beta subunit (also called ApoJalpha) corresponds to residues 23-227. The alpha subunit (also called ApoJbeta) corresponds to residues 228-449. Overexpression of Clusterin appears to be more common in late stages of mammary tumor progression. Clusterin markedly influences beta-Amyloid structure and neuritic toxicity in vivo and may influence Alzheimer's disease pathogenesis.

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: CLU