Synthetic peptide, C-DENFHYKHDREG, from the internal region of human PPID.
Cyclophilin D (CyPD) is a member of the immunophilin family of proteins. Immunophilins are a family of soluble cytosolic receptors capable of binding to one of two major immunosuppressant agents-cyclosporin A (CsA) or FK506. Proteins which bind FK506 are termed FK506 Binding Proteins (FKBPs) and those which bind cyclosporin A are called cyclophilins (CyP). Immunophilins function as peptidyl prolyl cis-trans-isomerases (PPIase) whose activity is inhibited by their respective immunosuppressant compounds. As PPIases, immunophilins accelerate folding of some proteins both in vivo and in vitro by catalyzing slow steps in the initial folding and rearrangement of proline containing proteins. CyPD is located in the inter-membrane space of the mitochondria where it has been shown to be an integral member of the permeability transition pore complex which also includes the voltage dependent anion channel (VDAC) and the adenine nucleotide translocator (ANT). Applications: Suitable for use in ELISA and Western Blot. Other Applications not tested. Recommended Dilutions: ELISA: 1:64,000 Western Blot: 0.1-0.3ug/ml Optimal dilutions to be determined by the researcher Positive Control: Human Brain (Frontal Cortex) lysates Storage and Stability: May be stored at 4C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and store at -20C. Aliquots are stable for at least 12 monthsat -20C. 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. Peptide Blocking: Corresponding peptide is available for Peptide Blocking studies. See C8855-20B-P. Antibody is typically 0.5mg/ml and peptide is supplied as a 100ul pellet. When peptide is reconstituted in 200ul water, the concentration would also be 0.5mg/ml. To start, the best ratio would be 1:1 (molar excess of peptide relative to antibody when identical volumes are mixed). Mix equal volumes of peptide and antibody at the required dilution and leave at ambient temperature. It is best is to have two identical blots to be incubated with equal amount of antibody, but one with the antibody pre-adsorbed to the peptide for 20min. Then incubate and develop the two blots in parallel.
