Human SOX18 (Sex Determining Region Y Box Protein 18) CLIA Kit (HUES01133)

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HUES01133
€599

Description

ELISA Kit Technical ManualMSDS

Human SOX18 (Sex Determining Region Y Box Protein 18) 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 SOX18 . 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 SOX18 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 SOX18, 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 SOX18. The concentration of Human SOX18 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 range31.25-2000 pg/mL
Sensitivity18.75 pg/mL
Sample volume100uL
Sample typeSerum, plasma and other biological fluids
RepeatabilityCV < 15%

Specificity

This kit recognizes Human SOX18 in samples. No significant cross-reactivity or interference between Human SOX18 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
200051694
59748
5572155693
100022942
25006
2397423946
50011565
10499
1103211004
2505222
5366
52945266
1252653
2563
26082580
62.51338
1284
13111283
31.25643
705
674646
028
28
28--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human SOX18 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human SOX18 were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)102.41174.44900.4398.92160.50959.78
Standard deviation9.4313.0188.969.0813.53108.94
C V (%)9.217.469.889.188.4311.35

Recovery

The recovery of Human SOX18 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)89-10094
EDTA plasma (n=5)90-10597
Cell culture media (n=5)92-10599

Linearity

Samples were spiked with high concentrations of Human SOX18 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 (%)96-11399-11498-111
Average (%)104105105
1:4Range (%)100-114100-11989-104
Average (%)10710896
1:8Range (%)89-10085-10098-112
Average (%)9591106
1:16Range (%)103-11997-11086-98
Average (%)10910391

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 SOX18 (Sex Determining Region Y Box Protein 18) CLIA Kit (HUES01133) 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 SOX18 (Sex Determining Region Y Box Protein 18) CLIA Kit (HUES01133) Protein Information

UniProt Protein Function:SOX18: Binds to the consensus sequence 5'-AACAAAG-3' and is able to trans-activate transcription via this site. Defects in SOX18 are the cause of hypotrichosis- lymphedema-telangiectasia syndrome (HLTS).
UniProt Protein Details:

Protein type:DNA-binding; Transcription factor

Chromosomal Location of Human Ortholog: 20q13.33

Cellular Component: nuclear chromatin; nucleus

Biological Process: angiogenesis; blood vessel endothelial cell migration; embryonic heart tube development; hair cycle process; heart looping; lymphangiogenesis; mRNA transcription from RNA polymerase II promoter; negative regulation of transcription from RNA polymerase II promoter; negative regulation of transcription, DNA-dependent; positive regulation of transcription from RNA polymerase II promoter; positive regulation of transcription, DNA-dependent; vasculature development; vasculogenesis

Disease: Hypotrichosis-lymphedema-telangiectasia Syndrome; Hypotrichosis-lymphedema-telangiectasia-renal Defect Syndrome

NCBI Summary:This gene encodes a member of the SOX (SRY-related HMG-box) family of transcription factors involved in the regulation of embryonic development and in the determination of the cell fate. The encoded protein may act as a transcriptional regulator after forming a protein complex with other proteins. This protein plays a role in hair, blood vessel, and lymphatic vessel development. Mutations in this gene have been associated with recessive and dominant forms of hypotrichosis-lymphedema-telangiectasia. [provided by RefSeq, Jul 2008]
UniProt Code:P35713
NCBI GenInfo Identifier:12644232
NCBI Gene ID:54345
NCBI Accession:P35713.2
UniProt Secondary Accession:P35713,Q0VGA9, Q9NPH8,
UniProt Related Accession:P35713
Molecular Weight:40,891 Da
NCBI Full Name:Transcription factor SOX-18
NCBI Synonym Full Names:SRY-box 18
NCBI Official Symbol:SOX18  
NCBI Official Synonym Symbols:HLTS; HLTRS  
NCBI Protein Information:transcription factor SOX-18
UniProt Protein Name:Transcription factor SOX-18
Protein Family:Transcription factor
UniProt Gene Name:SOX18  
UniProt Entry Name:SOX18_HUMAN
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Additional Information

Product type:
ELISA
Reactivity:
Human
ELISA Type:
CLIA
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