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).Antigen Distribution
Réactivité croisée (Details)
Species reactivity (tested):Human. The antibody also stains a subpopulation of macrophages of Rhesus Monkey and Bovine tissues. It does not react with Swine tissues.
Purification
Affinity Chromatography on Protein G
Immunogène
Cultured Human monocytes. Remarks: The antigen is MRP8/14 (Calprotectin), the epitope involves parts of both subunits MRP8 and MRP14.
S100A8/A9
Reactivité: Humain
WB
Hôte: Lapin
Polyclonal
Cy3
Indications d'application
ELISA. Immunohistochemistry (Including Paraffin Sections after Predigestion with Trypsin orProteinase K): 1 μg/mLFixation: Acetone, Formalin/Paraffin. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
Protocole
Protocol with frozen, ice-cold acetone-fixed sections: The whole procedure is performed at room temperature1. Wash in PBS2. Block endogenous peroxidase3. Wash in PBS4. Block with 10% normal goat serum in PBS for 30min. in a humid chamber5. Incubate with primary antibody (dilution see datasheet) for 1h in a humid chamber6. Wash in PBS7. Incubate with secondary antibody (peroxidase-conjugated goat anti mouse IgG+IgM(H+L) minimal-cross reaction to human) for 1h in a humid chamber8. Wash in PBS9. Incubate with AEC substrate (3-amino-9-ethylcarbazol) for 12min. 10. Wash in PBS11. Counterstain with Mayer’s hemalumProtocol with formalin-fixed, paraffin-embedded sections: The whole procedure is performed at room temperature1. Deparaffinize and rehydrate tissue section2. Incubate the tissue section with proteinase K for 7min. 3. Wash in distilled water4. Block endogenous peroxidase5. Wash in PBS6. Block with 10% normal goat serum in PBS for 30min. in a humid chamber7. Incubate with primary antibody (dilution see datasheet) for 1h in a humid chamber8. Wash in PBS9. Incubate with secondary antibody (peroxidase-conjugated goat anti mouse IgG+IgM
Restrictions
For Research Use only
Concentration
1.8 mg/mL (by Absorbance at 280 nm)
Buffer
PBS, pH 7.2, 0.09 % Sodium Azide, None
Agent conservateur
Sodium azide
Précaution d'utilisation
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Conseil sur la manipulation
Avoid repeated freezing and thawing.
Stock
4 °C/-20 °C
Stockage commentaire
Prior to reconstitution store at 2-8 °C. Following reconstitution store undiluted at 2-8 °C for one month or (in aliquots) at -20 °C for longer.
Ghavami, Rashedi, Dattilo, Eshraghi, Chazin, Hashemi, Wesselborg, Kerkhoff, Los: "S100A8/A9 at low concentration promotes tumor cell growth via RAGE ligation and MAP kinase-dependent pathway." dans: Journal of leukocyte biology, Vol. 83, Issue 6, pp. 1484-92, (2008) (PubMed).
Altwegg, Neidhart, Hersberger, Müller, Eberli, Corti, Roffi, Sütsch, Gay, von Eckardstein, Wischnewsky, Lüscher, Maier: "Myeloid-related protein 8/14 complex is released by monocytes and granulocytes at the site of coronary occlusion: a novel, early, and sensitive marker of acute coronary syndromes." dans: European heart journal, Vol. 28, Issue 8, pp. 941-8, (2007) (PubMed).
Rosenberger, Thorey, Werner, Boukamp: "A novel regulator of telomerase. S100A8 mediates differentiation-dependent and calcium-induced inhibition of telomerase activity in the human epidermal keratinocyte line HaCaT." dans: The Journal of biological chemistry, Vol. 282, Issue 9, pp. 6126-35, (2007) (PubMed).
Antigène
S100A8/A9 Complex (Calprotectin) (S100A8/A9)
(S100 Calcium Binding Protein A8/A9 (S100A8/A9))
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: CAGA, CAGB, CFAG, Calgranulin A/B, Calprotectin, L1 Protein, MRP-14, MRP-8, P14, P8