Some important signaling proteins can be regulated by phosphorylation at a specific threonine followed by arginine or lysine at the +2 position.
For example, conventional PKC isozymes phosphorylate substrates containing serine or threonine with Arg or Lys at the-3,-2 and +2 positions (1–2).
c-Raf, a mitogen-activated protein kinase and the main effector recruited by GTP-bound Ras, is phosphorylated at Thr481 and Thr491 followed by Lys at the +2 position (3).
APP1 and ERF also contain a phosphorylated Thr followed by Arg or Lys at the +2 position.
Phosphorylation of these sites is important for their activities.
In order to determine the phosphorylation state of Thr in the Thr-X-Arg motif and to identify potential new phosphorylation sites in this motif, antibodies have been developed that recognize phosphorylated Thr followed by Arg or Lys at the +2 position.
Applications:Suitable for use in ELISA, Western Blot and Immunohistochemistry.
Other applications have not been tested.
Recommended Dilution:Western Blot: 1:1000Immunohistochemistry (Paraffin): 1:800 Requires HIER using sodium citrate buffer pH6.0 prior to staining
ELISA Peptide: 1:1000Optimal dilutions to be determined by the researcher.
Storage and Stability:May be stored at 4°C for short-term only.
Aliquot to avoid repeated freezing and thawing.
Store at -20°C.
Aliquots are stable for 12 months after receipt.
For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
仕様
Size:100ul
Host:rabbit
Source Antibody:human
Grade:Affinity Purified
Purity:Purified by Protein A and peptide affinity chromatography.
Form:Supplied as a liquid in 10mM HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol. No preservative added.
Specificity:Recognizes endogenous levels of proteins containing the phospho-Thr-X-Arg motif. Detects phosphorylated Thr followed by Arg or Lys at the +2 position, though its reactivity is lower for the Lys compared to Arg at the +2 position. Does not crossreact with nonphospho-Thr or phospho-Ser in the same motif. It recognizes phospho-Thr in the FFT*R motif in PKCbeta II but does not recognize phospho-Thr in other motifs that lack Lys or Arg at +2. Recognizes all species. Species Crossreactivity: human, mouse, rat and monkey
Isotype:IgG
Calc Applications Abbrev:E IHC WB
Calc Crossreactivity:Hu Mk Mo Rt
Immunogen:Synthetic peptide containing the phospho-Thr-X-Arg motif.