IL5 Antibody (OASA07901)

Catalog#: OASA07901

• Size: 0.1mg

• Description of Target: RABBIT ANTI HUMAN INTERLEUKIN-5
• Gene Symbol: IL5
• Alias Symbols: EDF; TRF; IL-5; IL5
• Protein Accession#: NP_000870.1
• Swissprot Id: P05113
• Host: Rabbit
• Protein Size (# AA): 134
• Specificity: IL-5
• Application: ELISA, WB
• Immunogen: The immunogen for anti-IL5 antibody: recombinant human IL-5
• Product Format: Purified IgG - lyophilised
• Isotype: Polyclonal IgG
• Applications Info: ELISA : This antibody may be used in an indirect ELISA or as a capture reagent in a sandwich ELISA with This product B as the detection antibody
  Preservative Stabilisers: None present
  
  Antiserum Preparation: Antisera to human IL-5 were raised by repeated immunisations of rabbits with highly purified antigen. Purified IgG prepared by affinity chromatography.
  Approx Protein Conc: IgG concentration 1.0mg/ml after reconstitution
  
  Buffer Solutions: Phosphate buffered saline pH7.4
• Key Reference: 1. Campbell, H.D. et al. (1987) Molecular cloning, nucleotide sequence, and expression of the gene encoding human Eosinophil Differentiation Factor (Interleukin 5). P.N.A.S. 84: 6629-6633.
  2. Wang, J. et al. (1998) Circulating, but not local lung, IL-5 is required for the development of antigen-induced airways eosinophilia. J. Clin. Invest. 102: 1132-1141.
• Reconstitution and Storage: Reconstitution: Reconstitute with 0.1ml distilled water. For long term storage the addition of 0.09% sodium azide is recommended. NB: For functional studies do not add azide.
  Storage: Prior to reconstitution store at +4^oC.
  After reconstitution store at -20^oC.
  Storage in frost-free freezers is not recommended. This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.

Product Protocol: IL5 antibody used to evaluate ultrastructural immunogold localization of interleukin 5 to the crystalloid core compartment of eosinophil secondary granules in patients with atopic asthma (OASA07901)

Specificity: IL-5
Product Page: IL5 antibody (OASA07901)
Antibody Pair Available: OASA07901 Capture OASA07903 Detection
Experiment Type: lmmunogold Labeling to Demonstrate IL-5

Protocol:
Grids were rinsed for 10 min in 0.14% glycine in 0.1 M PBS, pH 7.2. The following sequence of steps was used to stain the grids, by placing them into reagent drops: (a) 10% normal goat serum (NGS) in PBS, 30 min; (b) 1:l00 dilution of primary mouse anti-human IL5 monoclonal antibody (MAb 7) (Dr. L. A. McNamee, Glaxo, Greenford. Middlesex, UK) in PBS for 2 hr at RT (c) wash in 0.14% glycine in 0.1 M PBS, pH 7.2; (d) secondary gold-labeled antibody [ 1:20 dilution of 10-nm colloidal gold conjugated to goat anti mouse immunoglobulins (GAM IgGIM) (Aurion; Wageningen. the Netherlands) in 0.1 M PBS, pH 7.2, for 2 hr]; (e) wash in 0.14% glycine in 0.1 M PBS; (f) wash for 3 min in distilled water. Control sections were incubated (a) with GAM IgGIM with omission of the primary antibody and (b) with an irrelevant IgGl MAb PSAIER-PR8 (Dakopatts; Glostrup, Denmark) of the same isotype and concentration. After drying, all grids were stained with uranyl acetate and lead citrate. Transmission micrographs were made on a Zeiss electron microscope 902 operating at 80 kV.

Summary:
1. Immunogold labeling of IL-5 was detected within the crystalloid core compartment of more than 90% of the eosinophil secondary granules visualized in ultra-thin sections of several eosinophilic granulocytes in the bronchial mucosa of two atopic asthmatic patientsWhen matrix compartment was visible, it was not detectably labeled for IL-5. Background labeling was negligible. 2. Other eosinophil granule populations (i.e., primary and small granules), as well as lipid bodies, were not labeled for IL-5, nor was nuclear or cytoplasmic staining detected. No staining was observed in mast cells or in T-lymphocytes. 3. These results were confirmed by two other anti-human IL-5 antibodies [monoclonal mouse IgGl anti-human IL5 (#2374-01) and polyclonal rabbit anti-human IL-5 (#1722-011)) from Genzyme (Cambridge, MA).

References:
1: Möller GM, de Jong TA, Overbeek SE, van der Kwast TH, Postma DS, Hoogsteden HC. Ultrastructural immunogold localization of interleukin 5 to the crystalloid core compartment of eosinophil secondary granules in patients with atopic asthma. J Histochem Cytochem. 1996 Jan;44(1):67-9. PubMed PMID: 8543784.