Aminopeptidase PILS/ARTS1 Overexpression Lysate

Western Blot: Aminopeptidase PILS/ARTS1 Overexpression Lysate [NBP2-04355] - Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for ARTS1.

• Reactivity: Hu
• Applications: WB

Aminopeptidase PILS/ARTS1 Overexpression Lysate Summary

• Description:

  Aminopeptidase PILS/ARTS1 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC214469
  
  Accession#: NM_016442
  
  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: ERAP1

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: 107.7 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 Aminopeptidase PILS/ARTS1 Overexpression Lysate

• Adipocyte-derived leucine aminopeptidase
• A-LAP
• A-LAPALAP
• Aminopeptidase PILS
• ARTS1
• ARTS1aminopeptidase regulator of TNFR1 shedding
• ARTS-1PILSAP
• EC 3.4.11
• EC 3.4.11.-
• EC 3.4.11.1
• endoplasmic reticulum aminopeptidase 1
• endoplasmic reticulum aminopeptidase associated with antigen processing
• ERAAP1
• ERAP1
• KIAA0525APPILS
• PILS-AP
• PILS-APERAAP
• Puromycin-insensitive leucyl-specific aminopeptidase
• Type 1 tumor necrosis factor receptor shedding aminopeptidase regulator

Background

The endoplasmic reticulum (ER) aminopeptidase 1 (ERAP1) is a 120 kDa protein localized to the lumen of the ER, which removes NH2-terminal residues from many antigenic precursors for MHC class I peptide presentation. Peptides that are presented by MHC class I on the surface of a cell must be 8-11 residues long, and ERAP1 specifically trims peptides of 9 amino acids or more. ERAP1 is also induced by interferon-gamma. The gene encoding human ERAP1 maps to chromosome 5q15. ERAP1 has previously been characterized as adipocyte-derived leucine aminopeptidase (A-LAP), puromycin-insensitive leucine-specific aminopeptidase (PILS-AP) and aminopeptidase regulator of TNFR1 shedding (ARTS-1). A-LAP is thought to inactivate several bioactive peptides, including angiotensin II and, subsequently, may be involved in the regulation of blood pressure. PILS-AP is described as playing a role in angiogenesis by regulating the proliferation and migration of endothelial cells, and ARTS-1 is characterized as a TNFR1 binding protein that promotes TNFR1 shedding. Further research will be necessary to fully elucidate the functions of this protein.

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: ERAP1