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 |
|
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
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
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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