Rat PTGES (Prostaglandin E Synthase, Microsomal) CLIA Kit (RTES00476)

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RTES00476
€649

Description

ELISA Kit Technical ManualMSDS

Rat PTGES (Prostaglandin E Synthase, Microsomal) 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 Rat PTGES . 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 Rat PTGES 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 Rat PTGES, 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 Rat PTGES. The concentration of Rat PTGES in the samples can be calculated by comparing the RLU of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityRat
Detection methodChemiluminescence
Detection range31.25-2000 pg/mL
Sensitivity18.75 pg/mL
Sample volume100uL
Sample typeSerum, plasma and other biological fluids
RepeatabilityCV < 15%

Specificity

This kit recognizes Rat PTGES in samples. No significant cross-reactivity or interference between Rat PTGES 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
200051557
59885
5572155693
100022104
25844
2397423946
50011410
10654
1103211004
2504821
5767
52945266
1252767
2449
26082580
62.51352
1270
13111283
31.25656
692
674646
027
29
28--

Precision

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

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

  Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)97.17293.69839.29106.50289.20758.69
Standard deviation10.0134.7162.0212.9721.0246.51
C V (%)10.3011.827.3912.187.276.13

Recovery

The recovery of Rat PTGES 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)94-107100
EDTA plasma (n=5)95-106100
Cell culture media (n=5)86-9891

Linearity

Samples were spiked with high concentrations of Rat PTGES 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 (%)92-10686-9987-101
Average (%)1009293
1:4Range (%)88-10288-99102-117
Average (%)9593108
1:8Range (%)91-10797-11291-103
Average (%)9810496
1:16Range (%)101-11593-10596-112
Average (%)10799104

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

Rat PTGES (Prostaglandin E Synthase, Microsomal) CLIA Kit (RTES00476) 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.

Rat PTGES (Prostaglandin E Synthase, Microsomal) CLIA Kit (RTES00476) Protein Information

UniProt Protein Function:PTGES: Catalyzes the oxidoreduction of prostaglandin endoperoxide H2 (PGH2) to prostaglandin E2 (PGE2). Belongs to the MAPEG family.
UniProt Protein Details:

Protein type:EC 5.3.99.3; Nuclear envelope; Isomerase; Membrane protein, multi-pass; Membrane protein, integral; Lipid Metabolism - arachidonic acid

Chromosomal Location of Human Ortholog: 9q34.3

Cellular Component: endoplasmic reticulum membrane; membrane; nuclear envelope lumen; perinuclear region of cytoplasm; integral to membrane

Molecular Function:glutathione binding; prostaglandin-E synthase activity

Biological Process: chronic inflammatory response; response to retinoic acid; cyclooxygenase pathway; response to lipopolysaccharide; prostaglandin biosynthetic process; signal transduction; response to organic cyclic substance; prostaglandin metabolic process; negative regulation of cell proliferation; response to cytokine stimulus; arachidonic acid metabolic process; response to calcium ion; acute inflammatory response

NCBI Summary:The protein encoded by this gene is a glutathione-dependent prostaglandin E synthase. The expression of this gene has been shown to be induced by proinflammatory cytokine interleukin 1 beta (IL1B). Its expression can also be induced by tumor suppressor protein TP53, and may be involved in TP53 induced apoptosis. Knockout studies in mice suggest that this gene may contribute to the pathogenesis of collagen-induced arthritis and mediate acute pain during inflammatory responses. [provided by RefSeq, Jul 2008]
UniProt Code:O14684
NCBI GenInfo Identifier:4758910
NCBI Gene ID:9536
NCBI Accession:NP_004869.1
UniProt Secondary Accession:O14684,O14900, Q5SZC0,
UniProt Related Accession:O14684
Molecular Weight:17kd
NCBI Full Name:prostaglandin E synthase
NCBI Synonym Full Names:prostaglandin E synthase
NCBI Official Symbol:PTGES  
NCBI Official Synonym Symbols:PGES; MPGES; PIG12; PP102; PP1294; MGST-IV; MGST1L1; TP53I12; mPGES-1; MGST1-L1  
NCBI Protein Information:prostaglandin E synthase
UniProt Protein Name:Prostaglandin E synthase
UniProt Synonym Protein Names:Microsomal glutathione S-transferase 1-like 1; MGST1-L1; Microsomal prostaglandin E synthase 1; MPGES-1; p53-induced gene 12 protein
UniProt Gene Name:PTGES  
UniProt Entry Name:PTGES_HUMAN
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Additional Information

Product type:
ELISA
Reactivity:
Rat
ELISA Type:
CLIA
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