Cat. No.
MABL-2858
Application
IP, WB, ELISA, FC, IHC
Isotype
Engineer antibody
Species Reactivity
Human
Clone No.
5.5
From
Recombinant Antibody
Specificity
mAb 5.5 specifically recognises complexed MRP-8/MRP-14 via the hydrophobic residues in helix IV of MRP-8 (S100A8) exposed during heterodimer formation. mAb recognition of MRP-8 & MRP-14 is calcium dependent. 5.5 epitope is most likely to be formed by linear sequence Phe26-His27-Ala28 in calcium-binding loop I of MRP-8, and by Phe68, Leu72, Ile76 and Lys77 that lie along one face of helix IV.
Alternative Names
MRP-8; MRP8; MRP-14; MRP14; CF antigen; CFAG; L1 molecule; p8,14; Calprotectin
UniProt
P05109; P06702
Immunogen
BALB/c mice were immunized using cells from acute monocytic leukemia patients. The BALB/c spleen cell suspension was fused with Sp2/0-Ag14 cells, and mAb 5.5 was isolated from the hybridoma cells either by radioimmunoassay with 125-I-labelled F(ab')2 fragments of sheep anti-mouse Ig as second layer, or by indirect immunofluorescence using rhodamine-conjugated goat anti-mouse Ig as the second layer.
Application Notes
mAb 5.5 has been used in immunoprecipitation and western blot experiments for the characterisation of p8,14 identified in extracts of monocytes and neutrophils. 5.5 was also used in flow cytometric analysis to show that p8,14 is not a cell surface protein, and in immunohistochemical staining to show that p8,14 is present extracellularly (Edgeworth 1991).
Antibody First Published
Hogg et al. Monoclonal antibody 5.5 reacts with p8, 14, a myeloid molecule associated with some vascular endothelium 1989. Eur J Immunol. 19(6):1053-61 PMID:2666142
Note on publication
Describes the activation of Myeloid cells by inflammatory stimuli, resulting in the secretion of p8,14 onto the endothelium. 5.5 was used for the detection of p8,14 to suggest that this molecule is released by myeloid cells adhering onto the endothelium, rather than the endothelium itself in response to inflammatory stimuli.
Size
100 μg Purified antibody.
Concentration
1 mg/ml.
Purification
Protein A affinity purified
Buffer
PBS with 0.02% Proclin 300.
Storage Recommendation
Store at 4⁰C for up to 3 months. For longer storage, aliquot and store at - 20⁰C.
