BNP Overexpression Lysate

Western Blot: NPPB Overexpression Lysate [NBL1-13753] - Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for NPPB.

• Reactivity: Hu
• Applications: WB

BNP Overexpression Lysate Summary

• Description: BNP Transient Overexpression Lysate
  Expression Host: HEK293T
  
  Plasmid: RC206590
  
  Accession#: NM_002521
  
  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: NPPB

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: 11.9 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 BNP Overexpression Lysate

• BNF
• BNP
• brain type natriuretic peptide
• Gamma-brain natriuretic peptide
• natriuretic peptide B
• natriuretic peptide precursor B
• natriuretic peptides B
• natriuretic protein
• NPPB

Background

Brain natriuretic peptide (BNP) circulates in blood as a peptide hormone with natriuretic, vasodilatory and renin inhibitory properties. BNP is secreted predominantly by the left ventricular myocytes in response to volume expansion and pressure overload. BNP belongs to a family of structurally similar peptide hormones, which includes atrial natriuretic peptide (ANP), BNP, C-type natriuretic peptide (CNP) and urodilatin. These peptides are characterized by a common 17 amino acid ring structure with a disulfide bond between two cystein residues. This ring structure shows high homology between different natriuretic peptides (eleven of the 17 amino acid residues are homologous in the ring of each of the natriuretic peptides, see fig. 18). BNP is a 32 amino acid peptide with disulfide bond between the cystein residues Cys10 and Cys26. In earlier studies it has been demonstrated that BNP concentration in blood increases with the severity of congestive heart failure. Quantitative measurement of BNP in blood provides an objective indicator of congestive heart failure severity

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: NPPB