S100A9 ELISA Kit

Catalog No. ABIN2859311

Product Details S100A9 ELISA Kit

Target: S100A9 (S100 Calcium Binding Protein A9 (S100A9))

Binding Specificity: AA 2-113

Reactivity: Mouse

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 31.2-2000 pg/mL

Minimum Detection Limit: 31.2 pg/mL

Application: ELISA

Purpose: Sandwich High Sensitivity ELISA kit for Quantitative Detection of Mouse S100A9

Brand: PicoKine™

Sample Type: Cell Culture Supernatant, Serum, Plasma (heparin), Plasma (EDTA)

Analytical Method: Quantitative

Specificity: Expression system for standard: E.coli
Immunogen sequence: A2-K113

Cross-Reactivity (Details): There is no detectable cross-reactivity with other relevant proteins.

Sensitivity: <10pg/mL

Material not included: Microplate reader in standard size. Automated plate washer. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. Clean tubes and Eppendorf tubes. Washing buffer (neutral PBS or TBS). Preparation of 0.01M TBS: Add 1.2g Tris, 8.5g Nacl

Immunogen: Expression system for standard: E.coli
Immunogen sequence: A2-K113

Application Details

Application Notes: Before using Kit, spin tubes and bring down all components to bottom of tube. Duplicate well assay was recommended for both standard and sample testing.

Comment:

Sequence similarities: Belongs to the S-100 family.

Plate: Pre-coated

Protocol: mouse S100A9 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for S100A9 has been precoated onto 96-well plates. Standards(E.coli, A2-K113) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for S100A9 is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the mouse S100A9 amount of sample captured in plate.

Assay Procedure:

Aliquot 0.1 mL per well of the 2000pg/mL,1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.2pg/mL mouse S100A9 standard solutions into the precoated 96-well plate. Add 0.1 mL of the sample diluent buffer into the control well (Zero well). Add 0.1 mL of each properly diluted sample of mouse cell culture supernates, serum or plasma(heparin, EDTA) to each empty well. See "Sample Dilution Guideline" above for details. It is recommended that each mouse S100A9 standard solution and each sample be measured in duplicate.

Assay Precision:

• Sample 1: n=16, Mean(pg/ml): 390, Standard deviation: 18.33, CV(%): 4.7
• Sample 2: n=16, Mean(pg/ml): 578, Standard deviation: 23.7, CV(%): 4.1
• Sample 3: n=16, Mean(pg/ml): 1032, Standard deviation: 54.7, CV(%): 5.3,
• Sample 1: n=24, Mean(pg/ml): 445, Standard deviation: 24.92, CV(%): 5.6
• Sample 2: n=24, Mean(pg/ml): 617, Standard deviation: 30.23, CV(%): 4.9
• Sample 3: n=24, Mean(pg/ml): 1358, Standard deviation: 84.2, CV(%): 6.2

Restrictions: For Research Use only

Handling

Handling Advice: Avoid multiple freeze-thaw cycles.

Storage: -20 °C,4 °C

Storage Comment: Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles

Expiry Date: 12 months

Target Details for S100A9

Target: S100A9 (S100 Calcium Binding Protein A9 (S100A9))

Alternative Name: S100A9 (S100A9 Products)

Background:

Protein Function: S100A9 is a calcium- and zinc-binding protein which plays a prominent role in the regulation of inflammatory processes and immune response. It can induce neutrophil chemotaxis, adhesion, can increase the bactericidal activity of neutrophils by promoting phagocytosis via activation of SYK, PI3K/AKT, and ERK1/2 and can induce degranulation of neutrophils by a MAPK-dependent mechanism. Predominantly found as calprotectin (S100A8/A9) which has a wide plethora of intra- and extracellular functions. The intracellular functions include: facilitating leukocyte arachidonic acid trafficking and metabolism, modulation of the tubulin-dependent cytoskeleton during migration of phagocytes and activation of the neutrophilic NADPH-oxidase. Activates NADPH- oxidase by facilitating the enzyme complex assembly at the cell membrane, transferring arachidonic acid, an essential cofactor, to the enzyme complex and S100A8 contributes to the enzyme assembly by directly binding to NCF2/P67PHOX. The extracellular functions involve proinfammatory, antimicrobial, oxidant-scavenging and apoptosis-inducing activities. Its proinflammatory activity includes recruitment of leukocytes, promotion of cytokine and chemokine production, and regulation of leukocyte adhesion and migration. Acts as an alarmin or a danger associated molecular pattern (DAMP) molecule and stimulates innate immune cells via binding to pattern recognition receptors such as Toll-like receptor 4 (TLR4) and receptor for advanced glycation endproducts (AGER). Binding to TLR4 and AGER activates the MAP-kinase and NF- kappa-B signaling pathways resulting in the amplification of the proinflammatory cascade. Has antimicrobial activity towards bacteria and fungi and exerts its antimicrobial activity probably via chelation of Zn(2+) which is essential for microbial growth. Can induce cell death via autophagy and apoptosis and this occurs through the cross-talk of mitochondria and lysosomes via reactive oxygen species (ROS) and the process involves BNIP3. Can regulate neutrophil number and apoptosis by an anti-apoptotic effect, regulates cell survival via ITGAM/ITGB and TLR4 and a signaling mechanism involving MEK-ERK. Its role as an oxidant scavenger has a protective role in preventing exaggerated tissue damage by scavenging oxidants. The iNOS-S100A8/A9 transnitrosylase complex is proposed to direct selective inflammatory stimulus-dependent S- nitrosylation of multiple targets such as GAPDH, NXA5, EZR, MSN and VIM by recognizing a [IL]-x-C-x-x-[DE] motif (By similarity). .

Background: Protein S100A9 also known as migration inhibitory factor-related protein 14 (MRP-14) or calgranulin-B is a protein that is encoded by the S100A9 gene. S100A9 is a member of the S100 family of proteins containing 2 EF hand calcium-binding motifs. It is mapped to Chromosome 3 in mouse. 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. This protein may function in the inhibition of casein kinase. S100A9 complexes with S100A8, another member of the S100 family of calcium-modulated proteins regulates myeloid cell function by binding to Toll-like receptor-4 (TLR-4) and the receptor for advanced glycation end products.

Synonyms: Protein S100-A9,Calgranulin-B,Leukocyte L1 complex heavy chain,Migration inhibitory factor-related protein 14,MRP-14,p14,S100 calcium-binding protein A9,S100a9,Cagb, Mrp14,

Full Gene Name: Protein S100-A9

Cellular Localisation: Secreted . Cytoplasm. Cytoplasm, cytoskeleton. Cell membrane, Peripheral membrane protein . Predominantly localized in the cytoplasm. Upon elevation of the intracellular calcium level, translocated from the cytoplasm to the cytoskeleton and the cell membrane. Upon neutrophil activation or endothelial adhesion of monocytes, is secreted via a microtubule-mediated, alternative pathway.

Gene ID: 20202

UniProt: P31725

Pathways: Transition Metal Ion Homeostasis, Positive Regulation of Endopeptidase Activity, S100 Proteins