Myeloperoxidase/MPO Overexpression Lysate

Western Blot: Myeloperoxidase Overexpression Lysate (Adult Normal) [NBL1-13201] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Myeloperoxidase.

• Reactivity: Hu
• Applications: WB

Myeloperoxidase/MPO Overexpression Lysate Summary

• Description:

  Myeloperoxidase/MPO Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC216029
  
  Accession#: NM_000250
  
  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: MPO

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: 83.7 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 Myeloperoxidase/MPO Overexpression Lysate

• EC 1.11.1
• EC 1.11.1.7
• Myeloperoxidase

Background

FUNCTION: Part of the host defense system of polymorphonuclear leukocytes. It is responsible for microbicidal activity against a wide range of organisms. In the stimulated PMN, MPO catalyzes the production of hypohalous acids, primarily hypochlorous acid in physiologic situations, and other toxic intermediates that greatly enhance PMN microbicidal activity. MPO is an important marker for myeloid cells, from the promyelocyte stage and to the mature forms. CATALYTIC ACTIVITY: Donor + H2O2 = oxidized donor + 2 H2O. CATALYTIC ACTIVITY: Cl- + H2O2 = HOCl + 2 H2O. COFACTOR: Binds 1 calcium ion per heterodimer. COFACTOR: Binds 1 heme B (iron-protoporphyrin IX) group covalently per heterodimer. SUBUNIT: Tetramer of two light chains and two heavy chains. SUBCELLULAR LOCATION: Lysosome. ALTERNATIVE PRODUCTS: 3 named isoforms produced by alternative splicing. DISEASE: Defects in MPO are the cause of myeloperoxidase deficiency (MPD). MPD is an autosomal recessive defect that results in disseminated candidiasis. SIMILARITY: Belongs to the peroxidase family. XPO subfamily.

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: MPO