Mouse PMP3 (Peroxisomal Membrane Protein 3) CLIA Kit (MOES00471)

SKU:
MOES00471
Product type:
ELISA
Reactivity:
Mouse
ELISA Type:
CLIA
€649

Description

ELISA Kit Technical ManualMSDS

Mouse PMP3 (Peroxisomal Membrane Protein 3) 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 Mouse PMP3 . 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 Mouse PMP3 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 Mouse PMP3, 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 Mouse PMP3. The concentration of Mouse PMP3 in the samples can be calculated by comparing the RLU of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityMouse
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 Mouse PMP3 in samples. No significant cross-reactivity or interference between Mouse PMP3 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
200054271
57171
5572155693
100021980
25968
2397423946
50011402
10662
1103211004
2504840
5748
52945266
1252782
2434
26082580
62.51411
1211
13111283
31.25646
702
674646
028
28
28--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse PMP3 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse PMP3 were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)94.04186.49821.76101.25197.99770.81
Standard deviation9.6722.2190.3910.0022.0884.40
C V (%)10.2811.9111.009.8811.1510.95

Recovery

The recovery of Mouse PMP3 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)84-9891
EDTA plasma (n=5)88-10295
Cell culture media (n=5)88-10295

Linearity

Samples were spiked with high concentrations of Mouse PMP3 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 (%)88-101103-11792-104
Average (%)9511097
1:4Range (%)90-10296-111101-115
Average (%)95102109
1:8Range (%)99-11498-11194-107
Average (%)105106100
1:16Range (%)100-11688-10192-106
Average (%)1079598

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

Mouse PMP3 (Peroxisomal Membrane Protein 3) CLIA Kit (MOES00471) 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.

Mouse PMP3 (Peroxisomal Membrane Protein 3) CLIA Kit (MOES00471) Protein Information

UniProt Protein Function:PXMP3: Somewhat implicated in the biogenesis of peroxisomes. Defects in PEX2 are the cause of peroxisome biogenesis disorder complementation group 5 (PBD-CG5); also known as PBD-CGF. PBD refers to a group of peroxisomal disorders arising from a failure of protein import into the peroxisomal membrane or matrix. The PBD group is comprised of four disorders: Zellweger syndrome (ZWS), neonatal adrenoleukodystrophy (NALD), infantile Refsum disease (IRD), and classical rhizomelic chondrodysplasia punctata (RCDP). ZWS, NALD and IRD are distinct from RCDP and constitute a clinical continuum of overlapping phenotypes known as the Zellweger spectrum. The PBD group is genetically heterogeneous with at least 14 distinct genetic groups as concluded from complementation studies. Defects in PEX2 are a cause of Zellweger syndrome (ZWS). ZWS is a fatal peroxisome biogenesis disorder characterized by dysmorphic facial features, hepatomegaly, ocular abnormalities, renal cysts, hearing impairment, profound psychomotor retardation, severe hypotonia and neonatal seizures. Death occurs within the first year of life. Defects in PEX2 are a cause of infantile Refsum disease (IRD). IRD is a mild peroxisome biogenesis disorder (PBD). Clinical features include early onset, mental retardation, minor facial dysmorphism, retinopathy, sensorineural hearing deficit, hepatomegaly, osteoporosis, failure to thrive, and hypocholesterolemia. The biochemical abnormalities include accumulation of phytanic acid, very long chain fatty acids (VLCFA), di- and trihydroxycholestanoic acid and pipecolic acid. Belongs to the pex2/pex10/pex12 family.
UniProt Protein Details:

Protein type:Motility/polarity/chemotaxis; Membrane protein, multi-pass; Cell development/differentiation; Ubiquitin conjugating system; Membrane protein, integral

Cellular Component: peroxisomal membrane; integral to peroxisomal membrane; membrane; integral to membrane; peroxisome

Molecular Function:zinc ion binding; metal ion binding

Biological Process: nervous system development; bile acid biosynthetic process; cholesterol homeostasis; fatty acid beta-oxidation; protein destabilization; very-long-chain fatty acid metabolic process; peroxisome organization and biogenesis; negative regulation of fibroblast proliferation; regulation of cholesterol biosynthetic process; protein import into peroxisome matrix; neuron migration; negative regulation of transcription from RNA polymerase II promoter; negative regulation of epithelial cell proliferation

UniProt Code:P55098
NCBI GenInfo Identifier:254028168
NCBI Gene ID:19302
NCBI Accession:NP_001156773.1
UniProt Secondary Accession:P55098,O35467,
UniProt Related Accession:P55098
Molecular Weight:34,732 Da
NCBI Full Name:peroxisome biogenesis factor 2
NCBI Synonym Full Names:peroxisomal biogenesis factor 2
NCBI Official Symbol:Pex2  
NCBI Official Synonym Symbols:PAF-1; PMP35; Pxmp3; D3Ertd138e  
NCBI Protein Information:peroxisome biogenesis factor 2; peroxin-2; Zellweger syndrome homolog; peroxisome assembly factor 1; peroxisomal membrane protein 3, 35 kDa
UniProt Protein Name:Peroxisome biogenesis factor 2
UniProt Synonym Protein Names:Peroxin-2; Peroxisomal membrane protein 3; Peroxisome assembly factor 1
Protein Family:Peroxisome biogenesis factor
UniProt Gene Name:Pex2  
UniProt Entry Name:PEX2_MOUSE
View AllClose