Human S100A9 (S100 Calcium Binding Protein A9) CLIA Kit (HUES00742)

SKU:
HUES00742
Product Type:
CLIA Kit
Size:
96 Assays
Sensitivity:
0.09ng/mL
Range:
0.16-10ng/mL
ELISA Type:
Sandwich
Synonyms:
MRP14, 60B8AG, CAGB, CFAG, CGLB, L1AG, LIAG, MAC387, MIF, NIF, P14, Calgranulin-B
Reactivity:
Human
Tested Sample Types:
Serum, plasma and other biological fluids
€599

Description

ELISA Kit Technical ManualMSDS

Human S100A9 (S100 Calcium Binding Protein A9) 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 S100A9 . 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 S100A9 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 S100A9, 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 S100A9. The concentration of Human S100A9 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 S100A9 in samples. No significant cross-reactivity or interference between Human S100A9 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
1031130
33812
3247132447
514992
15948
1547015446
2.57713
7293
75037479
1.253331
3975
36533629
0.631831
1691
17611737
0.31878
770
824800
0.16335
379
357333
024
24
24--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human S100A9 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human S100A9 were tested on 3 different plates, 20 replicates in each plate.

Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (ng/mL)0.531.494.040.531.434.14
Standard deviation0.050.130.410.060.170.34
C V (%)9.438.7210.1511.3211.898.21

Recovery

The recovery of Human S100A9 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)94-10699
EDTA plasma (n=5)94-107102
Cell culture media (n=5)91-10899

Linearity

Samples were spiked with high concentrations of Human S100A9 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 (%)85-97103-12093-105
Average (%)9010999
1:4Range (%)102-12198-11495-109
Average (%)110104102
1:8Range (%)94-10696-11196-112
Average (%)100102104
1:16Range (%)98-11196-11199-117
Average (%)103101107

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 S100A9 (S100 Calcium Binding Protein A9) CLIA Kit (HUES00742) 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 S100A9 (S100 Calcium Binding Protein A9) CLIA Kit (HUES00742) Protein Information

UniProt Protein Function:S100A9: a calcium-binding regulatory protein of the S-100 family expressed by macrophages in acutely inflammated tissues and in chronic inflammations. May be an inhibitor of protein kinases. Also expressed in epithelial cells constitutively or induced during dermatoses. May interact with components of the intermediate filaments in monocytes and epithelial cells. Interacts with CEACAM3 in a calcium-dependent manner.
UniProt Protein Details:

Protein type:Calcium-binding

Chromosomal Location of Human Ortholog: 1q21

Cellular Component: cytoskeleton; cytosol; extracellular region; extracellular space; nucleus; plasma membrane

Molecular Function:arachidonic acid binding; calcium ion binding; microtubule binding; protein binding; RAGE receptor binding; signal transducer activity; zinc ion binding

Biological Process: activation of NF-kappaB transcription factor; autophagy; caspase activation; cell-cell signaling; chemokine production; cytokine production; defense response to bacterium; defense response to fungus; inflammatory response; leukocyte migration during inflammatory response; neutrophil chemotaxis; positive regulation of cell growth; positive regulation of inflammatory response; regulation of cytoskeleton organization and biogenesis; sequestering of zinc ion

NCBI Summary:The protein encoded by this gene is a member of the S100 family of proteins containing 2 EF-hand calcium-binding motifs. S100 proteins are localized in the cytoplasm and/or nucleus of a wide range of cells, and involved in the regulation of a number of cellular processes such as cell cycle progression and differentiation. S100 genes include at least 13 members which are located as a cluster on chromosome 1q21. This protein may function in the inhibition of casein kinase and altered expression of this protein is associated with the disease cystic fibrosis. This antimicrobial protein exhibits antifungal and antibacterial activity. [provided by RefSeq, Nov 2014]
UniProt Code:P06702
NCBI GenInfo Identifier:115444
NCBI Gene ID:6280
NCBI Accession:P06702.1
UniProt Secondary Accession:P06702,Q6FGA1, Q9NYM0, Q9UCJ1, D3DV36,
UniProt Related Accession:P06702
Molecular Weight:13,242 Da
NCBI Full Name:Protein S100-A9
NCBI Synonym Full Names:S100 calcium binding protein A9
NCBI Official Symbol:S100A9
NCBI Official Synonym Symbols:MIF; NIF; P14; CAGB; CFAG; CGLB; L1AG; LIAG; MRP14; 60B8AG; MAC387
NCBI Protein Information:protein S100-A9
UniProt Protein Name:Protein S100-A9
UniProt Synonym Protein Names:Calgranulin-B; Calprotectin L1H subunit; Leukocyte L1 complex heavy chain; Migration inhibitory factor-related protein 14; MRP-14; p14; S100 calcium-binding protein A9
Protein Family:Protein
UniProt Gene Name:S100A9
UniProt Entry Name:S10A9_HUMAN
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