MaxLight™650 is a new Far-IR stable dye conjugate comparable to Alexa Fluor™647, DyLight™649, Cy5™ and offers better labeling efficiency, brighter imaging and increased immunodetection.
Protein kinase R (PKR) is transcriptionally induced by interferon and activated by double-stranded RNA (dsRNA).
PKR inhibits translation initiation through phosphorylation of the alpha subunit of the initiation factor eIF2 (eIF2alpha) and also controls the activation of several transcription factors such as NF-kappaB, p53 and Stats.
In addition, PKR mediates apoptosis induced by many different stimuli, such as LPS, TNF-alpha, viral infection and serum starvation.
Activation of PKR by dsRNA results in PKR dimerization and autophosphorylation at Thr446 and Thr451 in the activation loop.
Substitution of Thr451 to alanine completely inactivates PKR, while a mutant with a Thr446 to alanine substitution was partially active.
PKR phosphorylates the a subunit of eukaryotic initiation factor-2 and inhibits protein synthesis.
It is an important component of the interferon response to virus infection.
Recently, it has been reported that PKR also phosphorylates human p53 on serine392.
Applications:Suitable for use in Western Blotting.
Other applications not tested.
Recommended Dilution:Western Blot: detects phospho-PKR in RIPA lysates from HeLa cells that hadbeen stimulated with IFN alpha overnight followed by treatment with calyculin A for 15 minutes
Beadlyte Peptide Specificity Assay: dilutions were incubated with a cocktail ofmicrospheres conjugated to either the phosphorylated or non phosphorylated peptides.
Treated HeLa (lane 1) or untreated HeLa (lane2) cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with antiphospho-PKR (0.5µg/ml).
Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system.
Arrow indicates phospho-PKR (68kD).
Optimal dilutions to be determined by the researcher.
Storage and Stability:May be stored at 4ﾡC for short-term only.
For long-term storage, store at -20ﾡC.
Aliquots are stable for at least 12 months at -20ﾡC.
For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Further dilutions can be made in assay buffer.
Note: Applications are based on unconjugated antibody.
仕様
Size：100ul
Host：rabbit
Source Antibody：human
Grade：Affinity Purified
Purity：Purified by Protein A affinity chromatography.
Form：Supplied as a liquid in PBS, pH 7.2. No preservative added. Labeled with MaxLight™650.
Specificity：Recognizes human Protein Kinase R phosphorylated at Thr446, Mr 68kD. A non-specific protein was also detected, Mr ~75kD, which is inhibited by the immunizing peptide.
Isotype：IgG
Calc Applications Abbrev：WB
Calc Crossreactivity：Hu
Immunogen：Synthetic peptide corresponding to amino acids surrounding phosphorylated Thr446 of human Protein Kinase R, conjugated to KLH.
