Human LOX (Lysyl Oxidase) CLIA Kit (HUES00149)

SKU:
HUES00149
Product Type:
CLIA Kit
Size:
96 Assays
Sensitivity:
0.09ng/mL
Range:
0.16-10ng/mL
ELISA Type:
Sandwich
Synonyms:
Protein-Lysine 6-Oxidase, MGC105112
Reactivity:
Human
Tested Sample Types:
Serum, plasma and other biological fluids
€599

Description

ELISA Kit Technical ManualMSDS

Human LOX (Lysyl Oxidase) 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 LOX . 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 LOX 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 LOX, 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 LOX. The concentration of Human LOX 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 range0.16-10 ng/mL
Sensitivity0.09 ng/mL
Sample volume100uL
Sample typeSerum, plasma and other biological fluids
RepeatabilityCV < 15%

Specificity

This kit recognizes Human LOX in samples. No significant cross-reactivity or interference between Human LOX 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
(ng/mL)
RLU Average Corrected
1030644
34298
3247132447
514617
16323
1547015446
2.58083
6923
75037479
1.253499
3807
36533629
0.631898
1624
17611737
0.31885
763
824800
0.16334
380
357333
024
24
24--

Precision

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

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

Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (ng/mL)0.561.503.840.541.523.98
Standard deviation0.070.140.430.040.170.26
C V (%)12.509.3311.207.4111.186.53

Recovery

The recovery of Human LOX 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)91-10396
EDTA plasma (n=5)101-116107
Cell culture media (n=5)86-9792

Linearity

Samples were spiked with high concentrations of Human LOX 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 (%)86-99102-11798-109
Average (%)91109103
1:4Range (%)96-11386-9888-102
Average (%)1049193
1:8Range (%)97-11389-102103-117
Average (%)10395109
1:16Range (%)99-11688-10198-112
Average (%)10694106

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 LOX (Lysyl Oxidase) CLIA Kit (HUES00149) 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 LOX (Lysyl Oxidase) CLIA Kit (HUES00149) Protein Information

UniProt Protein Function:LOX: Responsible for the post-translational oxidative deamination of peptidyl lysine residues in precursors to fibrous collagen and elastin. In addition to cross-linking of extracellular matrix proteins, may have a direct role in tumor suppression. Belongs to the lysyl oxidase family.
UniProt Protein Details:

Protein type:EC 1.4.3.13; Secreted; Secreted, signal peptide; Oxidoreductase

Chromosomal Location of Human Ortholog: 5q23.2

Cellular Component: extracellular region

Molecular Function:protein binding

Biological Process: extracellular matrix organization and biogenesis; protein modification process

Disease: Cutis Laxa, Autosomal Recessive, Type Ia

NCBI Summary:This gene encodes a member of the lysyl oxidase family of proteins. Alternative splicing results in multiple transcript variants, at least one of which encodes a preproprotein that is proteolytically processed to generate a regulatory propeptide and the mature enzyme. The copper-dependent amine oxidase activity of this enzyme functions in the crosslinking of collagens and elastin, while the propeptide may play a role in tumor suppression. In addition, defects in this gene have been linked with predisposition to thoracic aortic aneurysms and dissections. [provided by RefSeq, Jul 2016]
UniProt Code:P28300
NCBI GenInfo Identifier:417269
NCBI Gene ID:4015
NCBI Accession:P28300.2
UniProt Secondary Accession:P28300,Q5FWF0, B2R5Q3,
UniProt Related Accession:P28300
Molecular Weight:46,944 Da
NCBI Full Name:Protein-lysine 6-oxidase
NCBI Synonym Full Names:lysyl oxidase
NCBI Official Symbol:LOX
NCBI Protein Information:protein-lysine 6-oxidase
UniProt Protein Name:Protein-lysine 6-oxidase
UniProt Synonym Protein Names:Lysyl oxidase
Protein Family:Linoleate 9/13-lipoxygenase
UniProt Gene Name:LOX
UniProt Entry Name:LYOX_HUMAN
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