Calprotectin (S100A8+A9 complex) Antibody (Mouse mAb) [H13D24]

Application: Reactivity: Human

Usage Information

Dilution
WB1:1000 IHC1:100 FCM1:10

Application
WB, FCM

Reactivity
Human

Source
Mouse Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW
13 kDa

Positive Control	Human tonsil tissue; Human peripheral blood mononuclear cells
Negative Control

Datasheet & SDS

Biological Description

Specificity
Calprotectin (S100A8+A9 complex) Antibody (Mouse mAb) [H13D24] detects endogenous levels of total S100A8/S100A9 calprotectin complex.

Clone
H13D24

Synonym(s)
CAGB, CFAG, MRP14, S100A9, Protein S100-A9, Calgranulin-B, Calprotectin L1H subunit, Leukocyte L1 complex heavy chain, Migration inhibitory factor-related protein 14, S100 calcium-binding protein A9, MRP-14, p14

Background
Calprotectin is the heterodimeric complex formed by S100A8 and S100A9, two calcium- and zinc-binding proteins of the S100 family that are among the most abundant cytoplasmic proteins in neutrophils and monocytes, and the complex operates through distinct intracellular and extracellular mechanisms rather than a single unified pathway. Intracellularly, calprotectin facilitates arachidonic acid trafficking within leukocytes and directly promotes assembly of the NADPH-oxidase enzyme complex at the plasma membrane, with S100A8 contributing to this assembly by binding directly to the cytosolic oxidase subunit NCF2/p67phox and by transferring arachidonic acid, an essential cofactor, to the assembling enzyme complex, coupling calprotectin's calcium-sensing function directly to activation of the neutrophil respiratory burst. Once released extracellularly from activated phagocytes, calprotectin functions as an alarmin, engaging the pattern-recognition receptors Toll-like receptor 4 and the receptor for advanced glycation end-products; mechanistic dissection of the TLR4 interaction identifies S100A8 as the specifically active component, which interacts directly with the TLR4-MD2 receptor complex and drives intracellular translocation of the adaptor protein MyD88 together with activation of interleukin-1 receptor-associated kinase-1, culminating in NF-kB activation and elevated tumor necrosis factor-alpha expression. Genetic loss of the S100A8-S100A9 complex protects mice from endotoxin-induced lethal shock and from Escherichia coli-induced abdominal sepsis, directly demonstrating that calprotectin release amplifies rather than merely accompanies the endotoxin-triggered inflammatory response of phagocytes during systemic infection. Beyond TLR4, calprotectin engagement of pattern-recognition receptors activates parallel MAP-kinase signaling that converges on and amplifies the same NF-kB-driven pro-inflammatory cascade, producing recruitment of additional leukocytes and increased cytokine and chemokine output at sites of tissue injury or infection. Calprotectin additionally exerts antimicrobial activity through chelation of zinc, a metal essential for microbial growth, and can trigger cell death in target cells through a pathway linking mitochondrial and lysosomal reactive oxygen species generation to the protein BNIP3, indicating that calprotectin's antimicrobial and apoptosis-inducing functions operate through mechanisms distinct from its TLR4-NF-kB pro-inflammatory signaling.

Background

Calprotectin is the heterodimeric complex formed by S100A8 and S100A9, two calcium- and zinc-binding proteins of the S100 family that are among the most abundant cytoplasmic proteins in neutrophils and monocytes, and the complex operates through distinct intracellular and extracellular mechanisms rather than a single unified pathway. Intracellularly, calprotectin facilitates arachidonic acid trafficking within leukocytes and directly promotes assembly of the NADPH-oxidase enzyme complex at the plasma membrane, with S100A8 contributing to this assembly by binding directly to the cytosolic oxidase subunit NCF2/p67phox and by transferring arachidonic acid, an essential cofactor, to the assembling enzyme complex, coupling calprotectin's calcium-sensing function directly to activation of the neutrophil respiratory burst. Once released extracellularly from activated phagocytes, calprotectin functions as an alarmin, engaging the pattern-recognition receptors Toll-like receptor 4 and the receptor for advanced glycation end-products; mechanistic dissection of the TLR4 interaction identifies S100A8 as the specifically active component, which interacts directly with the TLR4-MD2 receptor complex and drives intracellular translocation of the adaptor protein MyD88 together with activation of interleukin-1 receptor-associated kinase-1, culminating in NF-kB activation and elevated tumor necrosis factor-alpha expression. Genetic loss of the S100A8-S100A9 complex protects mice from endotoxin-induced lethal shock and from Escherichia coli-induced abdominal sepsis, directly demonstrating that calprotectin release amplifies rather than merely accompanies the endotoxin-triggered inflammatory response of phagocytes during systemic infection. Beyond TLR4, calprotectin engagement of pattern-recognition receptors activates parallel MAP-kinase signaling that converges on and amplifies the same NF-kB-driven pro-inflammatory cascade, producing recruitment of additional leukocytes and increased cytokine and chemokine output at sites of tissue injury or infection. Calprotectin additionally exerts antimicrobial activity through chelation of zinc, a metal essential for microbial growth, and can trigger cell death in target cells through a pathway linking mitochondrial and lysosomal reactive oxygen species generation to the protein BNIP3, indicating that calprotectin's antimicrobial and apoptosis-inducing functions operate through mechanisms distinct from its TLR4-NF-kB pro-inflammatory signaling.

References
https://pubmed.ncbi.nlm.nih.gov/12084065/ https://pubmed.ncbi.nlm.nih.gov/17767165/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%