| Lipoprotein-associated phospholipase A2 (Lp-PLA2) also known as platelet-activating factor acetylhydrolase (PAF-AH) is a phospholipase A2 enzyme that in humans is encoded by the PLA2G7 gene. Lp-PLA2 is a 45kD protein of 441 amino acids. It is one of several PAF acetylhydrolases. In the blood it travels mainly with low-density lipoprotein (LDL). Less than 20% is associated with high-density lipoprotein HDL. It is an enzyme produced by inflammatory cells and hydrolyzes oxidized phospholipids in LDL. Lp-PLA2 is platelet-activating factor (PAF) acetylhydrolase (EC 3.1.1.47), a secreted enzyme that catalyzes the degradation of PAF to inactive products by hydrolysis of the acetyl group at the sn-2 position, producing the biologically inactive products LYSO-PAF and acetate.[4] Lp-PLA2 is involved in the development of atherosclerosis, an observation that has prompted interest as a possible therapeutic target (see, e.g. the investigational drug Darapladib). In human atherosclerotic lesions, 2 main sources of Lp-PLA2 can be identified, including that which is brought into the intima bound to LDL (from the circulation), and that which is synthesized de novo by plaque inflammatory cells (macrophages, T cells, mast cells). Recombinant protein corresponding to aa22-441 with an N-Terminal His-Tag from human LpPLA2, expressed in E. coli (Q13093). Amino Acid Sequence: FDWQYINPV AHMKSSAWVN KIQVLMAAAS FGQTKIPRGN GPYSVGCTDL MFDHTNKGTF LRLYYPSQDN DRLDTLWIPN KEYFWGLSKF LGTHWLMGNI LRLLFGSMTT PANWNSPLRP GEKYPLVVFS HGLGAFRTLY SAIGIDLASH GFIVAAVEHR DRSASATYYF KDQSAAEIGD KSWLYLRTLK QEEETHIRNE QVRQRAKECS QALSLILDID HGKPVKNALD LKFDMEQLKD SIDREKIAVI GHSFGGATVI QTLSEDQRFR CGIALDAWMF PLGDEVYSRI PQPLFFINSE YFQYPANIIK MKKCYSPDKE RKMITIRGSV HQNFADFTFA TGKIIGHMLK LKGDIDSNVA IDLSNKASLA FLQKHLGLHK DFDQWDCLIE GDDENLIPGT NINTTNQHIM LQNSSGIEKY N Predicted Molecular Weight: 50.0kD Isoelectric Point: 7.1 Subcellular Location: Secreted, extracellular space Applications: Suitable for use in ELISA, Western Blot, Immunoprecipitation and SDS-PAGE. Other applications not tested. Recommended Dilutions: Optimal dilutions to be determined by the researcher. Included Protein Molecular Weight Marker: 168631: Unstained Protein Molecular Weight Marker, 10-70kD Suppled as a liquid in 62.5mM Tris-H3PO4, pH 7.5, 1mM EDTA, 2% SDS, 100mM DTT, 1mM sodium azide, 0.01% bromo-phenol blue, 33% glycerol. Ready to use, no need to heat, dilute or add reducing agents before use. Protein Bands: 10kD, 14kD, 18kD, 22kD, 26kD, 33kD, 44kD and 70kD. Double Intesity Bands: The 26kD, 18kD and 10kD bands are at double intensity to make location and size approximation of proteins easier. Usage: Allow the marker to reach room temperature and mix thoroughly before use to ensure that any solids that may have precipitated at -20C will go back into solution. Do not boil! Load the following volumes on SDS-PAGE gel: Mini gels: 3-5ul/well Large gel: 7ul/well The loading volumes listed above are recommended for gels with thickness of 0.75-1mm. The loading volume should be doubled for 1.5mm thick gels. Store at -20C Stable for 6 months after receipt. Storage and Stability: Lyophilized powder may be stored at -70C. Stable for 6 months after receipt. Reconstitute with sterile ddH2O. Aliquot to avoid repeated freezing and thawing. Store at -70C. 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: Thermal stability is described by the loss rate of the target protein. The loss rate was determined by accelerated thermal degradation by incubating the protein at 37C for 48h, with no obvious degradation or precipitation observed. Protein loss is <5% within the expiration date under appropriate storage conditions. |
