Calprotectin ELISA Kit

INTENDED USE

Human calprotectin ELISA kit can measure concentrations of S100A8/S100A9 heterodimer (calprotectin, S100 calcium binding protein, A8/S100 calcium binding protein A9) present is plasma, serum, biological fluids or tissue homogenate samples. This assay has a minimum sensitivity detection limit of 0.375 ng/ml.

INTRODUCTION

Calprotectin is abundantly found in neutrophils, it comprises nearly 60% of soluble proteins in the cytoplasm of these immune cells. It belongs to the larger S100 family of EF-hand calcium-binding proteins but has unique functions attributable to its heterodimeric structure of S100A8 and S100A9 subunits.

Released by activated phagocytes at sites of tissue injury and infection, calprotectin acts as a danger-associated molecular pattern or alarmin to signal tissue stress. By competing with pathogenic microbes for zinc and manganese ions via its zinc-binding His-X-His sites, it exhibits potent antimicrobial properties against bacteria and fungi. Defensins enhance this biostatic action. Additionally, extracellular calprotectin promotes inflammation by interacting with cell surface receptors like TLR4, RAGE, and EMMPRIN. This triggers the production of proinflammatory cytokines including IL-6 and TNFα.

Inside the cell, calprotectin regulates microtubule stability following stimuli such as RAS activation or extracellular calcium influx. Through reversible interactions with tubulins mediated by calcium levels, it plays a crucial role in microtubule dynamics and cytoskeletal rearrangements that govern cell motility and division. Calprotectin also shows promise as a molecular imaging biomarker in metastasis and cancer diagnosis as S100A8 and S100A9 are overexpressed by many tumor types.

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

• One 96-Well Plate: Pre-coated with anti-calprotectin antibody.
• Standards: Lyophilized recombinant.
• Sample/Standard Dilution Buffer.
• Biotinylated-labelled Antibody.
• Antibody Dilution Buffer.
• HRP-Streptavidin Conjugate (SABC).
• SABC Dilution Buffer.
• TMB Substrate.
• Wash Buffer (25x).
• Plate Sealer.
• Product Instructions.

TYPICAL RESULTS

For this calprotectin ELISA kit ELISA kit, it is recommended that a standard curve is generated for each assay carried out.

Standard Curve: 0, 0.625, 1.25, 2.5, 5, 10, 20, 40 ng/ml.
Reactivity: Human
Sensitivity: 0.375 ng/ml
Range: 0.625 – 40 ng/ml
Principle: Sandwich, Double Antibody
Application: Research Use Only.

ASSAY CHARACTERISTICS

– Specificity: Highly specific for calprotectin, no cross reactivity or interference between calprotectin and analogues was detected.
– Recovery: Serum (88 – 99%), EDTA Plasma (86 – 102%), Heparin Plasma (89 – 103%).
– Linearity: Serum (85 – 103%), EDTA Plasma (82 – 100%), Heparin Plasma (81 – 98%).
– Precison Intra-Assay: CV < 8%
– Precison Inter-Assay: CV < 10%

REFERENCES

1. Calprotectin: from biomarker to biological function. Gut. (2021) 70 (10): 1978-1988. Jukic A., et al.
2. Calprotectin in rheumatic diseases. Exp Biol Med (Maywood). (2017) 242 (8): 859-873. Ometto F., et al.
3. Calprotectin and cardiovascular events. A narrative review. Clin Biochem. (2014) 47 (12): 996-1001. Montagnana M., et al.
4. Formation of Calprotectin-Derived Peptides in the Airways of Children with Cystic Fibrosis. J Immunol. (2022) 208 (4): 979-990. Edwards T.S., et al.

ADDITIONAL INFORMATION

• Full Name: Calprotectin ELISA Kit
• Reactivity: Human
• Sample Type: Plasma, Tissue Homogenates, Serum, Biological Fluids
• Sensitivity: 0.375 ng/ml

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