Introduction
Kallistatin, also known as SERPINA4, acts as an inhibitor of tissue kallikrein, effectively hindering both its amidolytic and kininogenase functions in humans. This inhibitory action is achieved through the formation of a highly stable, equimolar complex between kallistatin and the enzyme. The complex is resistant to heat and SDS. Notably, the interaction leads to the cleavage of kallistatin at a specific reactive site by tissue kallikrein, resulting in the generation of a small C-terminal fragment.
Description
Recombinant human SERPINA4, expressed in HEK cells, is a single-chain polypeptide that has undergone glycosylation. It encompasses amino acids Gln21 to Pro427, resulting in a protein with 417 amino acids. With a predicted molecular weight of 47.7 kDa, this recombinant SERPINA4 is further modified by the addition of a 10-amino acid histidine tag at its C-terminus.
Physical Appearance
White powder, lyophilized and sterile-filtered.
Formulation
The SERPINA4 protein underwent a 0.4 μm filtration process before being lyophilized. The lyophilization was performed from a solution containing 0.5 mg/ml SERPINA4 in phosphate-buffered saline (pH 7.4) supplemented with 5% (w/v) Trehalose.
Solubility
To generate a working stock solution, it is advisable to add deionized water to the lyophilized pellet, aiming for a concentration of roughly 0.5 mg/ml. Allow adequate time for the pellet to completely dissolve. It's essential to note that SERPINA4 is not sterile in its lyophilized form. Prior to using it for cell culture applications, ensure sterility by filtering the reconstituted protein through an appropriate sterile filter.
Stability
For long-term storage, maintain the lyophilized SERPINA4 protein at -20°C. After reconstitution, it's recommended to aliquot the protein solution to minimize freeze-thaw cycles. The reconstituted protein demonstrates stability at 4°C for a limited duration; storage at this temperature for two weeks has shown no discernible changes in the protein.
Purity
Purity levels exceed 95.0%, as determined by SDS-PAGE analysis.
Synonyms
Serpin Peptidase Inhibitor, Clade A (Alpha-1 Antiproteinase, Antitrypsin) Member 4, PI4, KST, Serine (Or Cysteine) Proteinase Inhibitor, Clade A (Alpha-1 Antiproteinase, Antitrypsin), Member 4, Peptidase Inhibitor 4, Kallikrein Inhibitor, Serpin A4, PI-4, Protease Inhibitor 4 (Kallistatin), Kallistatin, KLST, KAL, SERPINA4.
Amino Acid Sequence
QLHVEHDGES CSNSSHQQIL ETGEGSPSLK IAPANADFAF RFYYLIASET PGKNIFFSPL SISAAYAMLS LGACSHSRSQ ILEGLGFNLT ELSESDVHRG FQHLLHTLNL PGHGLETRVG SALFLSHNLK FLAKFLNDTM AVYEAKLFHT NFYDTVGTIQ LINDHVKKET RGKIVDLVSE LKKDVLMVLV NYIYFKALWE KPFISSRTTP KDFYVDENTT VRVPMMLQDQ EHHWYLHDRY LPCSVLRMDY KGDATVFFIL PNQGKMREIE EVLTPEMLMR WNNLLRKRNF YKKLELHLPK FSISGSYVLD QILPRLGFTD LFSKWADLSG ITKQQKLEAS KSFHKATLDV DEAGTEAAAA TSFAIKFFSA QTNRHILRFN RPFLVVIFST STQSVLFLGK VVDPTKPHHH HHHHHHH.