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Serpin E1/PAI-1 Overexpression Lysate

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Western Blot: PAI1/Serpine1 Overexpression Lysate (Native) [NBP2-09008] - PAI1/Serpine1 Overexpression Lysate (Native) Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression ...read more

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

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Serpin E1/PAI-1 Overexpression Lysate Summary

Description

Serpin E1/PAI-1 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC202085

Accession#: NM_000602

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
SERPINE1

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
42.7 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 Serpin E1/PAI-1 Overexpression Lysate

  • Endothelial plasminogen activator inhibitor
  • Nexin
  • PAI1
  • PAI-1
  • PAI1PAI-1
  • PAISerpin E1
  • PLANH1
  • PLANH1plasminogen activator inhibitor 1
  • serine (or cysteine) proteinase inhibitor, clade E (nexin, plasminogenactivator inhibitor type 1), member 1
  • Serpin E1
  • serpin peptidase inhibitor, clade E (nexin, plasminogen activator inhibitortype 1), member 1

Background

PAI-1 and PAI-2 (for plasminogen activator inhibitor-1 and -2) are members of the serpin serine proteinase inhibitor family. PAI-1 and PAI-2 have been shown to regulate uPA (urokinase-type plasminogen activator) and tPA (tissue plasminogen activator), resulting in the inhibition of proteolytic activity. Members of the serpin family generally complex with their target proteinases, then disassociate slowly into cleaved species that fold into stable inactive forms. PAI-1 can fold into the inactive state without cleavage, resulting in the latent form of PAI-1. Activity can be restored to the latent form of PAI-1 through denaturation and renaturation. PAI-2 occurs in secreted and cytosolic forms through facultative polypeptide translocation. uPA is a serine proteinase that is a member of the trypsin family. It is responsible for the cleavage of plasminogen at the Arg-Val bond to produce plasmin. uPA consists of two chains designated A and B. The A chain can be cleaved, resulting in low and high molecular mass forms of uPA.

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.

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Bioinformatics

Gene Symbol SERPINE1