PLVAP Overexpression Lysate

Western Blot: PLVAP Overexpression Lysate (Adult Normal) [NBP2-06137] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for PLVAP.

• Reactivity: Hu
• Applications: WB

PLVAP Overexpression Lysate Summary

• Description:

  PLVAP Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC208914
  
  Accession#: NM_031310
  
  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: PLVAP

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: 50.4 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 PLVAP Overexpression Lysate

• FELS
• FELSfenestrated-endothelial linked structure protein; PV-1 protein
• Fenestrated endothelial-linked structure protein
• gp68
• plasmalemma vesicle associated protein
• PLVAP
• PV1
• PV-1
• PV1Plasmalemma vesicle protein 1
• PV-1plasmalemma vesicle-associated protein

Background

PLVAP (Plasmalemma vesicle associated protein) is a membrane associated protein of caveolae and is found in fenestral and stomatal diaphragms in fenestrated endothelia and transendothelial channels. Normally in skeletal and cardiac muscle, PLVAP expression is limited to small arterioles and venules; however, under conditions of inflammation, it can be induced on previously non expressing vessels in cardiac muscle. In the central nervous system (CNS), the panendothelial cell antigen expression is developmentally regulated. During embryonic development, the antigen is found on brain vasculature up to day 16 of gestation, after which it disappears. The cessation of PLVAP expression in the CNS may be associated with the establishment of the blood brain barrier, which begins on day 16 of gestation. In the adult mouse, inflammation in the CNS can lead to re expression of the panendothelial cell antigen. This antibody was raised by a genetic immunization technique. Genetic immunization can be used to generate antibodies by directly delivering antigen-coding DNA into the animal, rather than injecting a protein or peptide (Tang et al. PubMed: 1545867; Chambers and Johnston PubMed: 12910245; Barry and Johnston PubMed: 9234514). The animal`s cells produce the protein, which stimulates the animal`s immune system to produce antibodies against that particular protein. A vector coding for a partial fusion protein was used for genetic immunisation of a mouse and the resulting serum was tested in Western blot against an E.coli lysate containing that partial fusion protein. Genetic immunization offers enormous advantages over the traditional protein-based immunization method. DNA is faster, cheaper and easier to produce and can be produced by standard techniques readily amenable to automation. Furthermore, the antibodies generated by genetic immunization are usually of superior quality with regard to specificity, affinity and recognizing the native 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: PLVAP