sPLA₂ (human Type IIA) ELISA Kit

Name: sPLA₂ (human Type IIA) ELISA Kit
SKU: DEIA5043
Rating: 5 (1 reviews)

Regulatory status: For research use only, not for use in diagnostic procedures.

COA

SDS

Datasheet

Summary

Sample

Plasma, synovial fluid, and other sample matrices

Species Reactivity

Human

Detection Method

sELISA

Intended Use

sPLA₂ (human Type IIA) ELISA Kit is a sandwich assay that can be used for quantification of sPLA₂ in plasma, synovial fluid, and other sample matrices. The ELISA has a range from 31.3-2,000 pg/ml, with a limit of quantification of 31.3 pg/ml.

Contents of Kit

1.Anti-sPLA₂ (human Type IIA) ELISA Strip Plate, 1 plate
2.Anti-sPLA₂ (human Type IIA) HRP Conjugate, 1 vial/1.5 ml
3.sPLA₂ (recombinant human Type IIA) ELISA Standard, 2 vials/0.5 ml
4.ELISA Buffer Concentrate (10X), 2 vials/10 ml
5.Wash Buffer Concentrate (400X), 1 vial/5 ml
6.Polysorbate 20, 1 vial/3 ml
7.96-Well Cover Sheet, 1 cover
8.TMB Substrate Solution, 1 vial/12 ml
9.HRP Stop Solution, 1 vial/12 ml

Storage

This kit will perform as specified if stored as directed at -20°C and used before the expiration date indicated on the outside of the box.

Precision

Intra-assay precision was determined by analyzing at least 20 replicates of 3 matrix controls (human plasma samples) in a single assay.

Inter-assay precision was determined by analyzing replicates of 3 matrix controls (human plasma samples) in separate assays spanning across several days.

Detection Range

31.3-2,000 pg/ml

Sensitivity

The Lower Limit of Quantification (LLOQ) of the assay is 31.3 pg/ml.
The Lower Limit of Detection (LLOD) of the assay is 9.3 pg/ml.

General Description

Phospholipases A2 (PLA₂s) make up a superfamily of enzymes that hydrolyze the ester bond of phosphoglycerides at the sn-2 position to release free fatty acid and lysophospholipids. This superfamily is divided into three groups by molecular weight and Ca²⁺-dependence. Secreted PLA₂s (sPLA₂s) are small ~14 kDa enzymes that require millimolar concentrations of Ca²⁺. This family of enzymes is further subdivided based on the number of intramolecular disulfide bonds, and the presence or absence of a C-terminal extension and a three amino acid elapid loop (residues 54-56).¹
sPLA₂s are found in the venom of certain snakes (Types IA, IIA, and IIB from vipers, cobras, rattlesnakes, and kraits), in pancreatic juices (Type IB), in rat and murine testes (Type IIC), in placenta, synovial fluids and platelets (Type IIA), and in heart, placenta, lung, mast cells, and P388D1 macrophages (Type V).² Until 1994, the sPLA₂ responsible for the release of arachidonic acid in inflammation was believed to be Type IIA. Although recently discovered isoforms of PLA₂ clearly contribute to the release of arachidonic acid, sPLA₂ (Type IIA) continues to be a protein of interest in the field of inflammation.
This ELISA is specific for Type IIA sPLA₂, and does not cross react with Type I, Type IV, Type V PLA₂, inflammatory mediators such as tumor necrosis factor, interleukin-1, or platelet-activating factor.

Standard Curve

The standard curve presented is an example of the data typically obtained with this kit; however, your results will not be identical to these. You must run a new standard curve. Do not use the data below to determine the values of your samples. Your results could differ substantially.

Citations

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Documents

pdf COA

pdf SDS

pdf Datasheet

QA & Reviews

Customer Reviews

References

MicroRNA-877-5p is involved in the trovafloxacin-induced liver injury

TOXICOLOGY LETTERS

Authors: Mitsugi, Ryo; Itoh, Tomoo; Fujiwara, Ryoichi

Abstract

Trovafloxacin develops severe hepatotoxicity; however, the underlying mechanism of the trovafloxacin-induced liver injury has not been cleared. It has been shown that microRNAs (miRNAs) can be involved in the development of drug-induced liver injuries. We performed a miRNA microarray analysis to identify hepatic miRNAs that were induced or reduced by trovafloxacin in mice. It was demonstrated that miR-877-5p was the most increased miRNA in the mouse liver 24 h after the trovafloxacin administration. To investigate the role of miR-877-5p in the liver, we established miR-877-5p-overexpressed HepG2 cells. Microarray analysis detected altered expressions in 2077 (> 2-fold) and 1547 (<0.5-fold) genes in the miR-877-5p overexpressing cells compared to the mock cells. Especially, SLCO4C1, PEPCK, MT1M, HIST1H2BM, LGI1, and PLA2G2A were markedly increased or decreased in the miR-877-5p overexpressing cells. We conducted a correlation analysis between the expression levels of miR-877-5p and the six genes in eight miR-877-5p stably-expressed clones. It was shown that the PEPCK expression levels were correlated with miR-877-5p expression levels. PEPCK is associated with development of apoptotic cell death; therefore, the increased miR-877-5p-induced PEPCK can be a trigger that is involved in the development of trovafloxacin-induced liver injury. (C) 2016 Elsevier Ireland Ltd. All rights reserved.

Surfactant protein B inhibits secretory phospholipase A(2) hydrolysis of surfactant phospholipids

AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY

Authors: Hite, R. Duncan; Grier, Bonnie L.; Waite, B. Moseley; Veldhuizen, Ruud A.; Possmayer, Fred; Yao, Li-Juan; Seeds, Michael C.

Abstract

Hite RD, Grier BL, Waite BM, Veldhuizen RA, Possmayer F, Yao L, Seeds MC. Surfactant protein B inhibits secretory phospholipase A(2) hydrolysis of surfactant phospholipids. Am J Physiol Lung Cell Mol Physiol 302: L257-L265, 2012. First published October 28, 2011; doi:10.1152/ajplung.00054.2011.-Hydrolysis of surfactant phospholipids (PL) by secretory phospholipases A(2) (sPLA(2)) contributes to surfactant damage in inflammatory airway diseases such as acute lung injury/acute respiratory distress syndrome. We and others have reported that each sPLA(2) exhibits specificity in hydrolyzing different PLs in pulmonary surfactant and that the presence of hydrophilic surfactant protein A (SP-A) alters sPLA(2)-mediated hydrolysis. This report tests the hypothesis that hydrophobic SP-B also inhibits sPLA(2)-mediated surfactant hydrolysis. Three surfactant preparations were used containing varied amounts of SP-B and radiolabeled tracers of phosphatidylcholine (PC) or phosphatidylglycerol (PG): 1) washed ovine surfactant (OS) (pre- and postorganic extraction) compared with Survanta (protein poor), 2) Survanta supplemented with purified bovine SP-B (1-5%, wt/wt), and 3) a mixture of dipalmitoylphosphatidylcholine (DPPC), 1-palmitoyl-2-oleoyl-phosphatidylcholine (POPC), and 1-palmitoyl-2-oleoyl-phosphatidylglycerol (POPG) (DPPC: POPC: POPG, 40: 40: 20) prepared as vesicles and monomolecular films in the presence or absence of SP-B. Hydrolysis of PG and PC by Group IB sPLA(2) (PLA2G1A) was significantly lower in the extracted OS, which contains SP-B, compared with Survanta (P = 0.005), which is SP-B poor. Hydrolysis of PG and PC in nonextracted OS, which contains all SPs, was lower than both Survanta and extracted OS. When Survanta was supplemented with 1% SP-B, PG and PC hydrolysis by PLA2G1B was significantly lower (P < 0.001) than in Survanta alone. When supplemented into pure lipid vesicles and monomolecular films composed of PG and PC mixtures, SP-B also inhibited hydrolysis by both PLA2G1B and Group IIA sPLA2 (PLA2G2A). In films, PLA2G1B hydrolyzed surfactant PL monolayers at surface pressures <= 30 mN/m (P < 0.01), and SP-B lowered the surface pressure range at which hydrolysis can occur. These results suggest the hydrophobic SP, SP-B, protects alveolar surfactant PL from hydrolysis mediated by multiple sPLA2 in both vesicles (alveolar subphase) and monomolecular films (air-liquid interface).