Certificate of Analysis and Data Sheet

Source:
E.Coli

Catalog No.
ENZ-325

Background:

Group IB secretory phospholipase A2 (sPLA2-IB) mediates cell proliferation, cell migration, hormone release and eicosanoid production via its receptor in peripheral tissues. In the CNS, high-affinity binding sites of sPLA2-IB have been documented. sPLA2-IB induced neuronal cell death in a concentrationdependent manner depending on PGD2 metabolites, especially Delta12-PGJ2 that might mediate sPLA2-IB-induced apoptosis. The secretory PLA2 (sPLA2) family, in which 10 isozymes have been identified, consists of lowmolecular weight, Ca2+-requiring secretory enzymes that have been implicated in a number of biological processes, such as modification of eicosanoid generation, inflammation, and host defense.

Description:

The Recombinant Human Secreted Phospholipase A2-IB is manufactured with N-terminal fusionf HisTag. The Recombinant Human Secreted Phospholipase A2-IB His-Tagged Fusion Protein is 16 kDa containing 126 amino acid residues of the human secreted phospholipase A2-IB and 16 additional amino acid residues - HisTag.

amino acid sequence:

MRGSHHHHHH GMASHMAVWQ FRKMIKCVIP GSDPFLEYNN YGCYCGLGGS GTPVDELDKC CQTHDNCYDQ AKKLDSCKFL LDNPYTHTYS YSCSGSAITC SSKNKECEAF ICNCDRNAAI CFSKAPYNKA HKNLDTKKYC QS

Physical Appearance:

Sterile Filtered lyophilized (freeze-dried) powder.

Formulation:

Sterile filtered and lyophilized from 0.5 mg/ml in 0.05M Acetate buffer pH4.

Specificity:

The amino acid sequence of the recombinant human Secreted Phospholipase A2-IB is 100% homologous to the amino acid sequence of the human Secreted Phospholipase A2-IB.

Purification Method:

Three-step procedure using affinity Ni-NTA chromatography and size exclusion chromatography before and after refolding.

Purity:

>95% (SDS -PAGE analyzed).

Reconstitution:

Add 0.2 ml of 0.1M Acetate buffer pH4 and let the lyophilized pellet dissolve completely. For conversion into higher pH value, we recommend intensive dilution by relevant buffer to a concentration of 10μg/ml. In higher concentrations the solubility of this antigen is limited.

Storage:

Store lyophilized protein at -20蚓. Aliquot the product after reconstitution to avoid repeated freezing/thawing cycles. Reconstituted protein can be stored at 4蚓 for a limited period of time; it does not show any change after two weeks at 4蚓.

Stability/Shelf Life:

The lyophilized protein remains stable until the expiry date when stored at -20蚓.

Applications:

Western blotting

References:

?Juhl K, Efanov AM, Olsen HL, Gromada J. Secretory phospholipase A2 is released from pancreatic beta-cells and stimulates insulin secretion via inhibition of ATP-dependent K+ channels. Biochem Biophys Res Commun. 2003 Oct 17; 310(2): 274-9.
?Beck GCh, Yard BA, Schulte J, Haak M, van Ackern K, van der Woude FJ, Kaszkin M. Secreted phospholipases A2 induce the expression of chemokines in microvascular endothelium. Biochem Biophys Res Commun. 2003 Jan 17; 300(3): 731-7.
?Hanasaki K, Arita H. Phospholipase A2 receptor: a regulator of biological functions of secretory phospholipase A2. Prostaglandins Other Lipid Mediat. 2002 Aug; 68-69: 71-82. Review.
?Silliman CC, Moore EE, Zallen G, Gonzalez R, Johnson JL, Elzi DJ, Meng X, Hanasaki K, Ishizaki J, Arita H, Ao L, England KM, Banerjee A. Presence of the M-type sPLA(2) receptor on neutrophils and its role in elastase release and adhesion. Am J Physiol Cell Physiol. 2002 Oct; 283(4): C1102- 13.
?Yagami T, Ueda K, Asakura K, Hayasaki-Kajiwara Y, Nakazato H, Sakaeda T, Hata S, Kuroda T, Takasu N, Hori Y. Group IB secretory phospholipase A2 induces neuronal cell death via apoptosis. J Neurochem. 2002 May; 81(3): 449-61.

Usage:

This material is offered by ProSpec-TechnoGene for research, laboratory or further evaluation purposes.

Gene:

Name:PLA2G1B
Synonyms:PLA2, PLA2A

Protein synonyms/aliases:

Phospholipase A2 precursor (EC 3.1.1.4)
    (Phosphatidylcholine 2-acylhydrolase)
    (Group IB phospholipase A2).

Protein Family:

Belongs to the phospholipase A2 family.

Protein Domains:

Domains:
•IPR013090   Phospholipase A2, active site
•IPR001211   Phospholipase A2

Related proteins:

•Recombinant Human Secreted Phospholipase A2-V
•Recombinant Human Secreted Phospholipase A2-IIA
•Recombinant Human Secreted Phospholipase A2-IID
•Recombinant Human Secreted Phospholipase A2-IIE
•Recombinant Human Secreted Phospholipase A2-X
•Recombinant Phospholipase A2 Bee Venom Protein
•Recombinant Human Secreted Phospholipase A2-XII

Protein links:

Precursor- Protein structure and amino acid sequence:

Latest Publications:

1. Group IB secretory phospholipase A2 stimulates leukotriene B4 production by a unique mechanism in human neutrophils.
Biochem Biophys Res Commun 2005 Aug 26;334(2): 500-8
2. Group IB secretory phospholipase A2 stimulates CXC chemokine ligand 8 production via ERK and NF-kappa B in human neutrophils.
J Immunol 2004 Nov 15;173(10):6433-9
3. The role of propeptide in the refolding of human group IB phospholipase A2.
Acta Biochim Biophys Sin (Shanghai) 2004 Sep;36(9):583-8
4. Group IB secretory phospholipase A2 promotes matrix metalloproteinase-2-mediated cell migration via the phosphatidylinositol 3-kinase and Akt pathway.
J Biol Chem 2004 Aug 27;279(35):36579-85
5. Comparative analysis of gene expression mechanisms between group IA and IB phospholipase A2 genes from sea snake Laticauda semifasciata.
Gene 2004 May 12;332:179-90
6. Group IB phospholipase A2 from Pseudonaja textilis.
Arch Biochem Biophys 2004 Jan 1;421(1):10-20

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中華民國95年06月06日更新