Monoclonal antibodies to lipocortin‐1 as probes for biological function

Abstract

We have developed two monoclonal antibodies to human lipocortin-1 (103 and 105) as reagents for quantitating the protein in biological systems and neutralizing its activity. Lipo 105 is a high affinity antibody that is functional in ELISA and Western blot formats. The antibody recognizes a site between amino acids 30 and 55 in the lipocortin-I sequence and can be used on native or denatured protein. Lipo 103 is an antibody that neutralizes the phospholipase A, inhibitory activity of lipocortin-1 by blocking binding of the protein to phospholipid surfaces. The antibody is specific for native human lipocortin-1. Lipo 103 was recently shown to block lipocortin-l-dependent differentiation of a squamous carcinoma cell line, demonstrating its usefulness as a probe for function.

Inhibitory effect of annexin I on synovial inflammation in rat adjuvant arthritis

“…Dexamethasone sodium phosphate (Sigma) was dissolved in ethanol and diluted in saline immediately prior to use. Specific anti-annexin I mAb (1-A and 1-B) were kindly provided by Dr. J. Browning (Biogen, Cambridge, MA) (11). The endotoxin content of all preparations was Ͻ0.05 units/ml, by Limulus assay (E-Toxate; Sigma).…”

Comparative analysis of Annexin A1-formyl peptide receptor 2/ALX expression in human leukocyte subsets

“…The anti-AnxA1 monoclonal antibody was previously described [33] while the monoclonal anti-FPR2/ALX was generated and purchased from Genovac. Allophycocyanin (APC)-anti-mouse IgG and human IgG (used to prevent nonspecific binding) were purchased from Biolegend.…”

Annexin 1 mediates the rapid anti-inflammatory effects of neutrophil-derived microparticles

“…Microparticles were stained with the following mAb at 10 g/mL: anti-AnxA1 (clone 1B 19 ); anti-CD62L (clone FMC46; Serotec, Oxford, United Kingdom); anti-CD62P (clone MCA796; Serotec); anti-CD14 (clone MCA1568; Serotec), or isotype control (MCA928 and MCA691; Serotec) for 1 hour at 4°C; a final staining with a rabbit anti-mouse IgG (STAR9B; Serotec) was conducted. In some cases FITC-labeled annexin V was added (10 L of manufacturer's solution; Serotec) to monitor exposure of phosphatidylserine on the outer side of the microparticles.…”