NCF4 Overexpression Lysate

Western Blot: NCF4 Overexpression Lysate (Adult Normal) [NBP2-09005] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for NCF4.

• Reactivity: Hu
• Applications: WB

NCF4 Overexpression Lysate Summary

• Description:

  NCF4 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC216094
  
  Accession#: NM_000631
  
  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: NCF4

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: 38.9 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 NCF4 Overexpression Lysate

• MGC3810
• NCF
• NCF-4
• neutrophil cytosol factor 4
• neutrophil cytosolic factor 4 (40kD)
• neutrophil cytosolic factor 4, 40kDa
• Neutrophil NADPH oxidase factor 4
• p40phox
• p40-phox
• SH3 and PX domain-containing protein 4
• SH3PXD4P40PHOX

Background

The NADPH oxidase generates superoxide in phagocytic cells. It is important for immunity and its deficiency leads to chronic granulomatous disease (CGD) (1). p40-phox is a newly isolated cytosolic component of the nicotinamide adenine dinucleotide phosphate (NADPH)-oxidase that copurifies with p67-phox. Preliminary evidence indicates that it is a component of the cytosolic complex (2). Maximal activation of NADPH oxidase requires formation of a complex between the p40-phox and p67-phox subunits via association of their PB1 domains. The crystal structure of the p40-phox/p67phox PB1 heterodimer reveals that both domains have a beta grasp topology and that they bind in a front-to-back arrangement through conserved electrostatic interactions between an acidic OPCA motif on p40-phox and basic residues in p67-phox (3).

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: NCF4