Prokineticin R1/PROKR1 Overexpression Lysate Summary
Description |
Prokineticin R1/PROKR1 Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC219745
Accession#: NM_138964
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 |
PROKR1 |
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 |
44.6 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 Prokineticin R1/PROKR1 Overexpression Lysate
Background
PKR (protein kinase, activatable by RNA) is a double-stranded (ds) RNA-activated protein kinase that plays a key role in the innate immunity response to viral infection in higher eukaryotes (reviewed in Langland et al, 2006). PKR contains an N-terminal dsRNA-binding domain that consists of consensus dsRNA-binding motifs and a C-terminal kinase domain containing conserved motifs for protein kinase activity. DsRNA is a well characterized danger signal that the cell uses recognize the presence of viral infection. PKR binds to dsRNA molecules during viral infection and triggers antiviral innate defense mechanisms including the induction of type I interferons and downregulation of gene expression. One of the most well characterized substrates of activated PKR is the eukaryotic tranlation initiation facter, eIF2. PKR phosphorylates eIF2 during virus infection, which ultimately leads to an inhibition in protein synthesis and block in viral replication. The key role played by PKR in innate responses to viral infections is underscored by the large number of DNA and RNA viruses, whose hosts range from insects to humans, that code for PKR inhibitors. PKR is also known as PRKR, EIF2AK1, and EIF2AK2. Recognizes PKR; human PKR is 942 amino acid 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.
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