MRP8/14 (S100A8/A9) Mouse Monoclonal Antibody [Clone ID: 27E10]

Specifications

Product Data
Clone Name	27E10
Applications	ELISA, IHC
Recommend Dilution	ELISA. Immunohistochemistry on frozen sections: 0.25 µg /ml Immunohistochemistry on Paraffin sections: 1 µg/ml (Proteinase K pretreatment for antigen retrieval is recommended). Fixation: Acetone, Formalin/Paraffin. Suggested positive control: Human tonsil.
Reactivity	Human
Host	Mouse
Clonality	Monoclonal
Immunogen	Cultured Human monocytes.
Specificity	This monoclonal 27E10 antibody is ideally suited for the detection of early inflammatory macrophages and thus for the classification of acute stage inflammation in tissue sections and in smears, the characterization of tumorous tissues and the in vitro monitoring of peripheral blood cell cultures. Clone 27E10 is unique in that it recognizes an epitope on the MRP8/14 heterocomplex that is not exposed on the individual subunits MRP8 or MRP14. The antibody reacts with Human subpopulations of macrophages, monocytes and granulocytes; peripheral blood monocytes carry the antigen extra- and intracellularly, neutrophils only intracellularly. Antigen Distribution  Isolated Cells: A subpopulation of monocytes and neutrophilic granulocytes are positive. Monocytes carry the antigen both on the surface and intracellularly, granulocytes exhibit it only intracellularly. Up to 80% of monocytes in early cultures (24-48h) are positive. No reaction has been seen with lymphocytes or platelets.  Tissue Sections: The antigen is found in macrophages in the red pulp of the spleen and in the liver; strongly expressed on macrophages in acute inflamed tissues (peridontitis, contact excema, urticaria, erythrodermia) where some endothelial and epidermal cells may also express this antigen; absent on normal resident mononuclear phagocytes in healthy tissues (skin, gut, thymus).
Formulation	PBS, pH 7.2 State: Purified State: Liquid purifed IgG fraction Stabilizer: None Preservative: 0.09% Sodium Azide
Concentration	lot specific
Purification	Affinity Chromatography on Protein G
Conjugation	Unconjugated
Storage Condition	Store undiluted at 2-8°C. DO NOT FREEZE!
Background	The antigen is produced by the heterocomplex formation of MRP8 (S100A8 or Calgranulin A) and MRP14 (S100A9 or Calgranulin B), two calcium binding proteins of the S 100 protein family.
Synonyms	Calgranulin A/B, Calprotectin, L1 Protein, CFAG, P8, P14, CAGA, CAGB, CFAG, MRP-8, MRP-14
Note	This antibody has been produced in vitro free of serum and is free of Bovine IgG Protocol: Protocol with frozen, ice-cold acetone-fixed sections:  The whole procedure is performed at room temperature 1. Wash in PBS 2. Block endogenous peroxidase 3. Wash in PBS 4. Block with 10% normal goat serum in PBS for 30min. in a humid chamber 5. Incubate with primary antibody (dilution see datasheet) for 1h in a humid chamber 6. Wash in PBS 7. Incubate with secondary antibody (peroxidase-conjugated goat anti mouse IgG+IgM (H+L) minimal-cross reaction to human) for 1h in a humid chamber 8. Wash in PBS 9. Incubate with AEC substrate (3-amino-9-ethylcarbazol) for 12min. 10. Wash in PBS 11. Counterstain with Mayer’s hemalum.  Protocol with formalin-fixed, paraffin-embedded sections: The whole procedure is performed at room temperature 1. Deparaffinize and rehydrate tissue section 2. Incubate the tissue section with proteinase K for 7min. 3. Wash in distilled water 4. Block endogenous peroxidase 5. Wash in PBS 6. Block with 10% normal goat serum in PBS for 30min. in a humid chamber 7. Incubate with primary antibody (dilution see datasheet) for 1h in a humid chamber 8. Wash in PBS 9. Incubate with secondary antibody (peroxidase-conjugated goat anti mouse IgG+IgM (H+L) minimal-cross reaction to human) for 1h in a humid chamber 10. Wash in PBS 11. Incubate with AEC substrate (3-amino-9-ethylcarbazol) for 12min. 12. Wash in PBS 13. Counterstain with Mayer’s
Reference Data

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