CloneID: 5.5
Antigen Long Description: 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.
Buffer Composition: PBS with 0.02% Proclin 300.
Chimeric Use Statement: This chimeric rabbit antibody was made using the variable domain sequences of the original Mouse IgG1 format, for improved compatibility with existing reagents, assays and techniques.
Available Custom Conjugation Options: AP, HRP, Fluorescein, APC, PE, Biotin Type A, Biotin Type B, Streptavidin, FluoroProbes 647H, Atto488, APC/Cy7, PE/Cy7
Uniprot Accession No.: P05109; P06702
Specificity Statement: 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.
Application Notes (Clone): 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).
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