Human PLA2G10 (Phospholipase A2, Group X) CLIA Kit (HUES00597)

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HUES00597
€599

Description

ELISA Kit Technical ManualMSDS

Human PLA2G10 (Phospholipase A2, Group X) CLIA Kit

This kit uses Sandwich-CLIA as the method. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Human PLA2G10 . Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human PLA2G10 and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human PLA2G10, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured spectrophotometrically by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Human PLA2G10. The concentration of Human PLA2G10 in the samples can be calculated by comparing the RLU of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityHuman
Detection methodChemiluminescence
Detection range7.81-500 pg/mL
Sensitivity4.69 pg/mL
Sample volume100uL
Sample typeSerum, plasma and other biological fluids
RepeatabilityCV < 15%

Specificity

This kit recognizes Human PLA2G10 in samples. No significant cross-reactivity or interference between Human PLA2G10 and analogues was observed.

Typical Data

As the RLU values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

Concentration
(pg/mL)
RLU Average Corrected
50049777
53297
5153751508
25020072
21924
2099820969
1259972
8820
93969367
62.54219
4805
45124483
31.252440
2158
22992270
15.631347
1153
12501221
7.81728
752
740711
028
30
29--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human PLA2G10 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human PLA2G10 were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)26.0576.92241.5523.4781.78264.40
Standard deviation3.178.9719.691.958.6716.55
C V (%)12.1711.668.158.3110.606.26

Recovery

The recovery of Human PLA2G10 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type Range (%) Average Recovery (%)
Serum (n=5)94-112102
EDTA plasma (n=5)86-9891
Cell culture media (n=5)88-10596

Linearity

Samples were spiked with high concentrations of Human PLA2G10 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

    Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
1:2Range (%)98-113101-11890-106
Average (%)10610998
1:4Range (%)87-10294-10986-98
Average (%)9410092
1:8Range (%)88-10095-11098-113
Average (%)94102105
1:16Range (%)93-10888-9994-105
Average (%)10093100

Kit Components & Storage

An unopened kit can be stored at 4'C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

Item Specifications Storage
Micro CLIA Plate(Dismountable)8 wells X12 strips-20'C, 6 months
Reference Standard2 vials
Concentrated Biotinylated Detection Ab (100X)1 vial, 120 uL
Concentrated HRP Conjugate (100X)1 vial, 120 uL-20'C(shading light), 6 months
Reference Standard & Sample Diluent1 vial, 20 mL4'C, 6 months
Biotinylated Detection Ab Diluent1 vial, 14 mL
HRP Conjugate Diluent1 vial, 14 mL
Concentrated Wash Buffer (25X)1 vial, 30 mL
Substrate Reagent A1 vial, 5 mL4'C (shading light)
Substrate Reagent B1 vial, 5 mL4'C (shading light)
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 copy

Human PLA2G10 (Phospholipase A2, Group X) CLIA Kit (HUES00597) Assay procedure

    1. Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
    2. Aliquot 100µl of standard solutions into the standard wells.
    3. Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
    4. Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids.) into test sample wells.
    5. Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
    6. Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix.Incubate for 1 hour at 37°C.
    7. Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
    8. Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
    9. Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
    10. Add 100µL of Substrate mixture solution to each well. Cover with a new plate seal andincubate for no more than 5 min at 37°C. Protect the plate from light.
    11. Determine the RLU value of each well immediately.

Human PLA2G10 (Phospholipase A2, Group X) CLIA Kit (HUES00597) Protein Information

UniProt Protein Function:PLA2G10: PA2 catalyzes the calcium-dependent hydrolysis of the 2- acyl groups in 3-sn-phosphoglycerides. Has a powerful potency for releasing arachidonic acid from cell membrane phospholipids. Prefers phosphatidylethanolamine and phosphatidylcholine liposomes to those of phosphatidylserine. Belongs to the phospholipase A2 family.
UniProt Protein Details:

Protein type:Cell development/differentiation; EC 3.1.1.4; Lipid Metabolism - alpha-linolenic acid; Lipid Metabolism - arachidonic acid; Lipid Metabolism - ether lipid; Lipid Metabolism - glycerophospholipid; Lipid Metabolism - linoleic acid; Motility/polarity/chemotaxis; Phospholipase; Secreted; Secreted, signal peptide

Chromosomal Location of Human Ortholog: 16p13.12

Cellular Component: extracellular region

Molecular Function:phospholipase A2 activity; phospholipase activity

Biological Process: axon guidance; cholesterol homeostasis; lysophospholipid transport; negative regulation of transcription factor activity; phosphatidic acid biosynthetic process; positive regulation of cellular protein metabolic process; positive regulation of prostaglandin secretion; regulation of macrophage activation

NCBI Summary:This gene encodes a member of the phospholipase A2 family of proteins. Alternative splicing results in multiple transcript variants, at least one of which encodes a preproprotein that is proteolytically processed to generate the mature enzyme. This calcium-dependent enzyme hydrolyzes glycerophospholipids to produce free fatty acids and lysophospholipids. In one example, this enzyme catalyzes the release of arachidonic acid from cell membrane phospholipids, thus playing a role in the production of various inflammatory lipid mediators, such as prostaglandins. The encoded protein may promote the survival of breast cancer cells through its role in lipid metabolism. [provided by RefSeq, Nov 2015]
UniProt Code:O15496
NCBI GenInfo Identifier:239938869
NCBI Gene ID:8399
NCBI Accession:O15496.3
UniProt Secondary Accession:O15496,Q14DU3, Q6NT23,
UniProt Related Accession:O15496
Molecular Weight:18kDa
NCBI Full Name:Group 10 secretory phospholipase A2
NCBI Synonym Full Names:phospholipase A2 group X
NCBI Official Symbol:PLA2G10  
NCBI Official Synonym Symbols:SPLA2; GXPLA2; GXSPLA2; sPLA2-X  
NCBI Protein Information:group 10 secretory phospholipase A2
UniProt Protein Name:Group 10 secretory phospholipase A2
UniProt Synonym Protein Names:Group X secretory phospholipase A2; GX sPLA2; sPLA2-X; Phosphatidylcholine 2-acylhydrolase 10
Protein Family:Group 10 secretory phospholipase
UniProt Gene Name:PLA2G10  
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Additional Information

Product type:
ELISA
Reactivity:
Human
ELISA Type:
CLIA
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