Leptin A Active Protein
Tilapia (Oreochromis niloticus) leptin A
Purity
The purity of recombinant tilapia leptins is greater than 95.0% as determined by:(a) Gel filtration analysis.
(b) Analysis by reducing and non-reducing SDS-PAGE gel.
Synonyms
Leptin A; Tilapia (Oreochromis niloticus) leptin A; Tilapia (Oreochromis niloticus) leptin A and B; Leptin A active protein
Host
E Coli
Purity/Purification
The purity of recombinant tilapia leptins is greater than 95.0% as determined by:
(a) Gel filtration analysis.
(b) Analysis by reducing and non-reducing SDS-PAGE gel.
(a) Gel filtration analysis.
(b) Analysis by reducing and non-reducing SDS-PAGE gel.
Form/Format
White lyophilized (freeze-dried) powder.
Sequence
The sequence of the first six N-terminal amino acids of recombinant Tilapia leptin A was determined and was found to be Ala-Pro-Leu-Pro-Val-Glu. The corresponding N-terminal sequence of Tilapia leptin B was Ala-Leu-Leu-Thr-Lys-Gly.
Solubility
It is recommended to reconstitute the lyophilized recombinant Tilapia leptins in sterile water or sterile 0.4% NaHCO3 adjusted to pH 8-9, not less than 100ug/ml, which can then be further diluted with other aqueous solutions.
Protein Content
Protein quantitation of recombinant fish leptin was carried out by UV spectroscopy at 280 nm using the absorbency value of 1.33 for 1 mg/ml (tLepA) and 0.19 for 1 mg/ml (tLepB) as the extinction coefficient for a 0.1% (1mg/ml) solution at pH 8.0. This value is calculated by the DNAman computer analysis program of protein sequences.
Dimers and Aggregates
The purified monomeric recombinant Tilapia leptin A (16K) consists of > 97% monomers as determined by gel-filtration chromatography. The covalent dimer of Tilapia leptin B contains less than 5% monomers. Tilapia leptin B consited of > 90% monomers.
Endotoxin
Less than 0.1 ng/ug (IEU/ug) of recombinant Tilapia leptins.
Formulation
The recombinant fish leptin was lyophilized from a concentrated (1.0 mg/ml) solution containing NaHCO3 at 1:2 salt:protein ratio.
Biological Activity
Both monomeric and dimeric Tilapia leptins were biologically active in promoting proliferation of BAF/3 cells stably transfected with the long form of human leptin receptor (hLepR), but their activity was four orders of magnitude lower than that of mammalian leptin. Furthermore, the Tilapia leptins were biologically active in promoting STATLUC activation in COS7 cells transfected with Tilapia leptin receptor but not in cells transfected with human leptin receptor. Tilapia Leptin A was more active than Tilapia Leptin B.
Preparation and Storage
Lyophilized recombinant tilapia leptins although stable at room temperature for several weeks, should be stored desiccated below -18 degree C. Upon reconstitution of recombinant tilapia leptins at > 0.1 mg/ml and up to 2 mg/ml and filter sterilization recombinant tilapia leptins can be stored at 4 degree C for several weeks. At lower concentration of recombinant tilapia leptins addition of a carrier protein (0.1% HSA or BSA) is suggested. Please prevent freeze-thaw cycles.
Related Product Information for Leptin A active protein
Fulllength cDNA encoding two leptin sequences (tLepA and tLepB) were identified in tilapia (Oreochromis niloticus). The cDNAs of tLepA and tLepB were 486 bp and 459 bp in length, encoding proteins of 161 aa and 152 aa, respectively. The tLepA and tLepBexpressing plasmids were transformed into E. coli and expressed as recombinant proteins upon induction with nalidixic acid, found almost entirely in insoluble inclusion bodies (IBs). The proteins were solubilized, refolded and purified to homogeneity by anionexchange chromatography. In the case of tLepA, the fraction eluted contained a mixture of monomers and dimers.
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Product Notes
The Leptin A (Catalog #AAA400122) is an Active Protein produced from E Coli and is intended for research purposes only. The product is available for immediate purchase. The amino acid sequence is listed below: The sequence of the first six N-terminal amino acids of recombinan t Tilapia leptin A was determined and was found to be Ala-Pro-Le u-Pro-Val- Glu. The correspond ing N-terminal sequence of Tilapia leptin B was Ala-Leu-Le u-Thr-Lys- Gly. It is sometimes possible for the material contained within the vial of "Leptin A, Active Protein" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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