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Monoclonal Antibodies

Anti-S100A8 & S100A9 [5.5]

Invented by Prof Nancy Hogg from Cancer Research UK London Research Institute: Lincoln's Inn Fields
Invented at Cancer Research UK London Research Institute: Lincoln's Inn Fields
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Adapted from Hessian PA, Fisher L. The heterodimeric complex of MRP-8 (S100A8) and MRP-14 (S100A9). Antibody recognition, epitope definition and the implications for structure. Eur J Biochem. 2001 Jan;268(2):353-63. doi: 10.1046/j.1432-1033.2001.01894.x. PMID: 11168370. Monoclonal antibodies 27E10 and 5.5 specifically recognize soluble MRP‐8/MRP‐14 complex. MRP‐8 or MRP‐14 were produced separately by in vitro transcription and translation or were cotranscribed/translated (MRP‐8/MRP‐14). To confirm the presence of protein products, radiolabelled MRP‐8 protein was immunoprecipitated with monoclonal antibody CF145 (A) and radiolabelled MRP‐14 with mAb MAC‐387 (B) in the presence of added 2 mm EGTA or 2 mm calcium as indicated. The relative areas to which MRP‐8 or MRP‐14 migrate following SDS/PAGE are shown by arrows. Equivalent aliquots of the same MRP‐8 or MRP‐14 monomer reactions were also immunoprecipitated with mAb 27E10 (C) or mAb 5.5 (D), also in the presence of added EGTA or calcium. No bands were visible. In vitro reactions where MRP‐8 and MRP‐14 were cotranscribed/translated (MRP‐8/MRP‐14) prior to immunoprecipitation with mAb MAC‐387 (B) mAb 27E10 (C) or mAb 5.5 (D) are also shown. The two proteins noncovalently associate only in the presence of added calcium to form epitopes recognized by mAbs 27E10 or 5.5. The complex is disrupted during SDS/PAGE and the separate monomers are distinguishable (arrowed). + and – signs indicate the presence or absence of critical components in each immunoprecipitation reaction. No reactivity with control mAbs FMC17 or D32 was observed (data not shown).
Adapted from Wang Y, Fang C, Gao H, Bilodeau ML, Zhang Z, Croce K, Liu S, Morooka T, Sakuma M, Nakajima K, Yoneda S, Shi C, Zidar D, Andre P, Stephens G, Silverstein RL, Hogg N, Schmaier AH, Simon DI. Platelet-derived S100 family member myeloid-related protein-14 regulates thrombosis. J Clin Invest. 2014 May;124(5):2160-71. doi: 10.1172/JCI70966. Epub 2014 Apr 1. PMID: 24691441; PMCID: PMC4001535. Platelets express MRP-8 and MRP-14.(A) MRP-8 (8 kDa) and MRP-14 (14 kDa) protein expression was examined in gel-filtered (lanes 1 and 2) and washed (lanes 3 and 4) WT and Mrp14–/– platelets. Platelets were lysed with SDS-PAGE–reduced sample buffer and then immunoblotted sequentially with anti–MRP-8, anti–MRP-14, and anti-tubulin antibodies. (B) Expression of MRP-8/14 in human platelets adherent to fibrinogen-coated coverslips was investigated using combined immunofluorescence with one antibody that recognizes the MRP-8/14 complex and another the platelet-specific marker GPIX. Robust MRP-8/14 staining was evident in GPIX-positive human platelets. Scale bars: 10 μm. (C) Platelet surface expression of MRP-14 in resting and thrombin-stimulated (FIIa-stimulated) platelets was determined by flow cytometry (mean fluorescence intensity; n = 4 per group). (D) MRP-8/14 and CD40L protein content in gel-filtered human platelets (mean ± SD, n = 5 normal human donors). (E) Immunoblot of plasma (18 μl per lane) from noninjured animals and from animals 10 minutes after photochemical carotid artery injury using anti–MRP-14 antibody. Equal loading was verified by IgG light chain. (F) Paired plasma and serum levels of MRP-8/14 (μg/ml) from 5 normal human donors.
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Catalogue Number 151056
Applications ELISA FACS IHC IP WB
Antigen/Gene or Protein Targets S100A8 (MRP-8) & S100A9 (MRP-14)
Reactivity Human
Relevance S100A8 (MRP-8) and S100A9 (MRP-14) are cytosolic calcium-binding proteins of 8kDa and 14kDa that form a heterodimer. S100A8 and S100A9 are expressed in secretory and inflamed keratinocytes, peripheral blood monocytes, neutrophils and has been described in platelets, dendritic cells and some T cell types. Expression is lost on tissue maturation of monocytes to macrophages. S100A9 may be associated with monocyte and neutrophil activation and the accumulation of these cells in inflammatory sites.
Host Mouse
Immunogen Acute monocytic leukaemia cells.
Positive Control This antibody was used to detect S100A8/S100A9 in neutrophil extracts (Hogg et al., 1989) This antibody was used to detect S100A8/S100A9 in Human monocyte and neutrophil extracts (Edgeworth et al., 1991)
Subclass IgG1
Recommended Growing Conditions DMEM + 5% FCS
Research Area Cell Cycle, Epigenetics & Nuclear Signalling

References

There are 12 reference entries for this reagent.

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References: 12 entries

Wen et al. 2018. Nat Med. 24(2):154-164. PMID: 29291352.

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Wang et al. 2014. J Clin Invest. 124(5):2160-71. PMID: 24691441.

View  

Ryckman et al. 2003. Arthritis Rheum. 48(8):2310-20. PMID: 12905486.

View  

Role of S100A8 and S100A9 in neutrophil recruitment in response to monosodium urate monohydrate crystals in the air-pouch model of acute gouty arthritis.

Europe PMC ID: 12905486

View  

Hessian et al. 2001. Eur J Biochem. 268(2):353-63. PMID: 11168370.

View  

Hessian et al. 2001. Eur J Biochem. 268(2):353-63. PMID: 11168370.

View  

The heterodimeric complex of MRP-8 (S100A8) and MRP-14 (S100A9). Antibody recognition, epitope definition and the implications for structure.

Europe PMC ID: 11168370

View  

Edgeworth et al. 1991. J Biol Chem. 266(12):7706-13. PMID: 2019594.

View  

Identification of p8,14 as a highly abundant heterodimeric calcium binding protein complex of myeloid cells.

Europe PMC ID: 2019594

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Hogg et al. 1989. Eur J Immunol. 19(6):1053-61. PMID: 2666142.

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Hogg et al. 1989. Eur J Immunol. 19(6):1053-61. PMID: 2666142.

View  

Monoclonal antibody 5.5 reacts with p8,14, a myeloid molecule associated with some vascular endothelium.

Europe PMC ID: 2666142

View  

Add a reference

References: 12 entries

Wen et al. 2018. Nat Med. 24(2):154-164. PMID: 29291352.

View  

Wang et al. 2014. J Clin Invest. 124(5):2160-71. PMID: 24691441.

View  

Ryckman et al. 2003. Arthritis Rheum. 48(8):2310-20. PMID: 12905486.

View  

Role of S100A8 and S100A9 in neutrophil recruitment in response to monosodium urate monohydrate crystals in the air-pouch model of acute gouty arthritis.

View  

Hessian et al. 2001. Eur J Biochem. 268(2):353-63. PMID: 11168370.

View  

Hessian et al. 2001. Eur J Biochem. 268(2):353-63. PMID: 11168370.

View  

The heterodimeric complex of MRP-8 (S100A8) and MRP-14 (S100A9). Antibody recognition, epitope definition and the implications for structure.

View  

Edgeworth et al. 1991. J Biol Chem. 266(12):7706-13. PMID: 2019594.

View  

Identification of p8,14 as a highly abundant heterodimeric calcium binding protein complex of myeloid cells.

View  

Hogg et al. 1989. Eur J Immunol. 19(6):1053-61. PMID: 2666142.

View  

Hogg et al. 1989. Eur J Immunol. 19(6):1053-61. PMID: 2666142.

View  

Monoclonal antibody 5.5 reacts with p8,14, a myeloid molecule associated with some vascular endothelium.

View  

Add a reference

Inventor Information