MRP8 (S100A8) Mouse Monoclonal Antibody [Clone ID: 8-5C2]

Specifications

Product Data
Clone Name	8-5C2
Applications	ELISA, IHC, WB
Recommend Dilution	ELISA. Immunohistochemistry on Frozen Sections: 1 µg/ml (1/200). Immunohistochemistry on Paraffin Sections: 4 µg/ml (1/20-1/50) (No pretreatment for antigen retrieval necessary). Microwave treatment in 0.01M Citrate, pH 6.0, may enhance the reactivity. Suggested Positive Control: Human tonsil. Has been described to work in Dot Blots and not in FACS.
Reactivity	Human
Host	Mouse
Clonality	Monoclonal
Immunogen	Cultured Human monocytes.
Specificity	This Monoclonal 8-5C2 antibody reacts with Human MRP8 in stimulated monocytes and macrophages in late phase or chronic inflammation. The antigen is MRP8, the epitope is suspected in the central portion of the peptide. Antigen Distribution on Isolated Cells: The antigen is found in granulocytes and monocytes but not in other blood cells. In cultured monocytes, maximum MRP8 is expressed after 3-4 days. Myeloid leukaemia stain positive. Antigen Distribution on Tissue Sections: MRP8 is found in a distinct subpopulation of inflammatory perivascular infiltrates of the myelo-monocytic lineage. Macrophages synthesise MRP8 increasingly during the late stages of inflammation. A low MRP8 (and high MRP- 14) expression by macrophages was also reported in granulomatous diseases such as tuberculosis and sarcoidis. In non-granulomatous chronic inflammatory diseases such as chronic rheumatoid arthritis or chronic rejection after allograft transplantation, MRP8 and MRP14 positive cells consist of different subpopulations.
Formulation	Stock Solution contains PBS, pH 7.2 State: Purified State: Lyophilized purified Ig fraction Stabilizer: 0.05% Kathon Preservative: 5 mg/ml BSA
Reconstitution Method	Restore with 0.5 ml distilled water.
Concentration	0.2 mg/ml (after reconstitution)
Purification	Affinity Chromatography
Conjugation	Unconjugated
Storage Condition	Store lyophilized at 2-8°C for 6 months or at -20°C long term. After reconstitution store the antibody undiluted at 2-8°C for one month  or (in aliquots) at -20°C long term. Avoid repeated freezing and thawing.
Gene Name	S100 calcium binding protein A8
Database Link	Entrez Gene 6279 Human UniProt ID P05109
Background	Monoclonal antibody 8-5C2 identifies MRP8 (also named S100A8 or Calgranulin A), the Ca2+-binding light subunit of the inflammatory L-1 protein complex. MRP8 forms Ca2+ dependent dimers or complexes with MRP14 (S100A9, Calgranulin B). It also forms disulfide-linked homodimers under the influence of hypochlorite, a process thought to abrogate the chemotactic property of MRP8. The antibody is useful in various immunological techniques. Histological and serological data indicate that MRP8 is associated with chronic stages of inflammatory diseases.
Synonyms	S100-A8, CAGA, MRP-8, CFAG
Note	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. Place slide in a cuvette with 250ml 0.01M citrate buffer, pH 6.0 3. Heat slide in a microwave oven for 2 x 7min. at 700Watt 4. Leave slide in the buffer for 20min for cooling 5. Wash in distilled water 6. Block endogenous peroxidase 7. Wash in PBS 8. Block with 10% normal goat serum in PBS for 30min. in a humid chamber 9. Incubate with primary antibody (dilution see datasheet) for 1h in a humid chamber 10. Wash in PBS 11. Incubate with secondary antibody (peroxidase-conjugated goat anti mouse IgG+IgM (H+L) minimal-cross reaction to human) for 1h in a humid chamber 12. Wash in PBS 13. Incubate with AEC substrate (3-amino-9-ethylcarbazol) for 12min. 14. Wash in PBS 15. Counterstain with Mayer’s hemalum
Reference Data

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