Clone S13.67 identifies MRP8: The antibody is useful in various immunological techniques. Histological and serological data indicate that MRP8 is associated with chronic stages of inflammatory diseases. This clone also stains cells in rat spleen, indiating significant cross reactivity with the corresponding Rat MRP8. Antigen Distribution on Isolated Cells: The antigen is found in granulocytes and monocytes. It is absent from other blood cells. In cultured monocytes, maximum MRP8 is expressed after 3-4 days. Antigen Distribution on Tissue Sections: In the tissue, MRP-8 is only found in a distinct subpopulation of inflammatory perivascular infiltrates of the myelo-monocytic lineage. Macrophages increasingly synthesise MRP-8 during the late stages of inflammation. A low MRP-8 (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 MRP8 and MRP14 positive cells consist of different subpopulations. During early inflammation endothelial cells are also positive with MRP8/MRP14.
Purification
Affinity Chromatography.
Immunogen
Cultured Human monocytes. Remarks: Antigen: MRP8. Epitope: Suspected in the N- or C-terminal Domain.
S100A8
Reactivity: Human, Mouse
ELISA, FACS, IF (cc), IF (p)
Host: Rabbit
Polyclonal
unconjugated
Application Notes
ELISA. Immunohistochemistry on Frozen Sections: 1-2 μg/mL (1/100-1/200). Immunohistochemistry on Paraffin Sections: 4 μg/mL (1/50). Pre-treatment for antigenretrieval not required. Has been described to work in FACS and Dot blots. Suggested Positive Control: Human tonsil. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
Protocol
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.
Restrictions
For Research Use only
Reconstitution
Restore by adding 0.5 mL distilled water.
Concentration
0.2 mg/mL
Buffer
Stock solution contains PBS, pH 7.2 with 5 mg/mL BSA as a stabilizer and 0.05 % (v/v) Kathon CG as preservative.
Preservative
Thimerosal (Merthiolate)
Precaution of Use
Do Not Use Sodium Azide as preservative.
Handling Advice
Avoid repeated freezing and thawing. Dilute only prior to immediate use
Storage
4 °C/-20 °C
Storage Comment
Store vial at 2-8 °C prior to restoration. For extended storage add glycerol to 50% and then aliquot contents and freeze at -20 °C or below. Centrifuge product if not completely clear after standing at room temperature. This antibody is stable for one month at 2-8 °C as an undiluted liquid.
Target
S100A8
(S100 Calcium Binding Protein A8 (S100A8))
Alternative Name
S100A8 / Calgranulin-A / MRP8
Background
MRP8 is the Ca2+-binding light subunit of Calprotectin. MRP8 forms Ca2+-dependent 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.Synonyms: CAGA, CFAG, Calprotectin L1L subunit, Cystic fibrosis antigen, Leukocyte L1 complex light chain, MRP-8, Migration inhibitory factor-related protein 8, S100 calcium-binding protein A8, S100-A8, Urinary stone protein band A