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Human S100A9 ELISA Kit

SKU:
HUFI01620
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P06702
Sensitivity:
0.469ng/ml
Range:
0.781-50ng/ml
ELISA Type:
Sandwich
Synonyms:
S100A9, S100 Calcium Binding Protein A9, Calgranulin B, MRP-14, CAGBMigRation inhibitory factor-related protein 14, Calgranulin-B, Calprotectin L1H subunit, CGLB, L1AG, LIAG, MRP14Leukocyte L1 complex heavy chain, NIF, P14, protein S100-A9
Reactivity:
Human
Research Area:
Cell Death
$599
Frequently bought together:

Description

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Human S100A9 ELISA Kit

The Human S100A9 ELISA Kit is specifically designed to accurately detect levels of S100A9 protein in human serum, plasma, and cell culture supernatants. This kit offers high sensitivity and specificity, providing reliable and reproducible results for a variety of research applications.S100A9 is a key protein involved in inflammatory responses and immune regulation. It has been linked to various conditions such as chronic inflammation, autoimmunity, and cancer, making it a valuable biomarker for studying these diseases and developing targeted treatments.

With the Human S100A9 ELISA Kit, researchers can confidently measure S100A9 levels in biological samples, advancing their understanding of disease mechanisms and potentially leading to the development of new therapeutic interventions.

Product Name:Human S100A9 ELISA Kit
Product Code:HUFI01620
Size:96 Assays
Alias:S100A9, S100 Calcium Binding Protein A9, Calgranulin B, MRP-14, CAGBMigRation inhibitory factor-related protein 14, Calgranulin-B, Calprotectin L1H subunit, CGLB, L1AG, LIAG, MRP14Leukocyte L1 complex heavy chain, NIF, P14, protein S100-A9
Detection method:Sandwich ELISA, Double Antibody
Application:This immunoassay kit allows for the in vitro quantitative determination of Human S100A9 concentrations in serum plasma and other biological fluids.
Sensitivity:0.469ng/ml
Range:0.781-50ng/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Human S100A9 and the recovery rates were calculated by comparing the measured value to the expected amount of Human S100A9 in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)90-10598
EDTA plasma(n=5)87-10396
UFH plasma(n=5)86-9793
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human S100A9 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)92-105%85-105%89-102%
EDTA plasma(n=5)85-96%84-100%86-100%
UFH plasma(n=5)80-93%82-97%85-100%
CV(%):Intra-Assay: CV<8%
Inter-Assay: CV<10%
ComponentQuantityStorage
ELISA Microplate (Dismountable)8×12 strips4°C for 6 months
Lyophilized Standard24°C/-20°C
Sample/Standard Dilution Buffer20ml4°C
Biotin-labeled Antibody(Concentrated)120ul4°C (Protect from light)
Antibody Dilution Buffer10ml4°C
HRP-Streptavidin Conjugate(SABC)120ul4°C (Protect from light)
SABC Dilution Buffer10ml4°C
TMB Substrate10ml4°C (Protect from light)
Stop Solution10ml4°C
Wash Buffer(25X)30ml4°C
Plate Sealer5-

Other materials and equipment required:

  • Microplate reader with 450 nm wavelength filter
  • Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
  • Incubator
  • Deionized or distilled water
  • Absorbent paper
  • Buffer resevoir
UniprotP06702
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

*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

Before adding to wells, equilibrate the SABC working solution and TMB substrate for at least 30 min at 37°C. When diluting samples and reagents, they must be mixed completely and evenly. It is recommended to plot a standard curve for each test.

StepProtocol
1.Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells!
2.Aliquot 0.1ml standard solutions into the standard wells.
3.Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well.
4.Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells.
5.Seal the plate with a cover and incubate at 37 °C for 90 min.
6.Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2.
7.Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall.
8.Seal the plate with a cover and incubate at 37°C for 60 min.
9.Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash.
10.Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min.
11.Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min.
12.Add 90 µl of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color.
13.Add 50 µl of Stop solution into each well and mix thoroughly. The color changes into yellow immediately.
14. Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution.

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Sample TypeProtocol
SerumIf using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

PlasmaCollect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit.
Urine & Cerebrospinal FluidCollect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.
Cell culture supernatantCollect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.
Cell lysatesSolubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Tissue homogenatesThe preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C.
Tissue lysatesRinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Breast MilkCollect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.

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