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Phosphate-buffered solution, pH 7.2, containing no preservative. 0.2 μm filter sterilized. Endotoxin level is < 0.1 EU/μg of the protein (< 0.01 ng/μg of the protein) as determined by the LAL test.
Preparation:
The LEAF™ (Low Endotoxin, Azide-Free) antibody was Purified by affinity chromatography.
Concentration:
1.0 mg/ml
Storage & Handling:
The antibody solution should be stored undiluted at 4 °C. This LEAF™ solution contains no preservative; handle under aseptic conditions.
Each lot of this antibody is quality control tested by ELISA assay. For ELISPOT capture applications, a concentration range of 2-8 µg/ml is recommended. For ELISA capture applications, a concentration range of 1-4 µg/ml is recommended. To obtain a linear standard curve, serial dilutions of mouse IL-5 recombinant protein ranging from 250 to 1 pg/ml are recommended for each ELISA plate when paired with TRFK4 (Cat. # 504402). To obtain a linear standard curve, serial dilutions of human IL-5 recombinant protein ranging from 250 to 1 pg/ml are recommended for each ELISA plate when paired with JES1-5A10 (Cat. # 501002). It is recommended that the reagent be titrated for optimal performance for each application.
COA:
Application Notes:
ELISA1,3,4 or ELISPOT2,6 Capture: The Purified TRFK5 antibody is useful as a capture antibody for a sandwich ELISA or ELISPOT assay, when used in conjunction with biotinylated TRFK4 antibody (Cat. No. 504402) as the detection antibody for mouse IL-5, or used in conjunction with biotinylated JES1-5A10 antibody (Cat. No. 501002/501005) as the detection antibody for human IL-5. The LEAF™ Purified antibody is suggested for ELISPOT capture. Flow Cytometry5: The fluorochrome-labeled TRFK5 antibody is useful for intracellular immunofluorescent staining and flow cytometric analysis to identify IL-5-producing cells within mixed cell populations. View intracellular cytokine staining protocol Neutralization1,9: The LEAF™ Purified antibody (Endotoxin <0.1 EU/μg, Azide-Free, 0.2 μm sterile-filtered) is recommended for neutralization of mouse and human IL-5 bioactivity (Cat. No. 504308). Additional reported applications (for the relevant formats) include: Western blotting, immunohistochemical staining of formalin-fixed paraffin-embedded tissue sections8 and paraformaldehyde-fixed, saponin-treated frozen tissue sections7, and immunocytochemistry. Note: For testing human IL-5 in serum or plasma, BioLegend's ELISA Max™ Sets (Cat. No. 430401 to 430406) are specially developed and recommended. For testing mouse IL-5 in serum, plasma or supernatant, BioLegend's ELISA Max™ Sets (Cat. No. 431201 to 431206) are specially developed and recommended.
Application References:
1. Abrams, J. 1995. Curr. Prot. Immunol. John Wiley and Sons, New York. Unit 6.20. 2. Klinman, D., et al. 1994. Curr. Prot. Immunol. John Wiley and Sons, New York. Unit 6.19. 3. Swain, S., et al. 1990. J. Immunol. 145:3796. 4. Abrams, J., et al. 1992. Immunol. Rev. 127:5. 5. Assenmacher, M., et al. 1994. Eur. J. Immunol. 24:1097. 6. Karulin, A., et al. 2000. J. Immunol. 164:1862. 7. Andersson, U., et al. 1999. Detection and quantification of gene expression. New York:Springer-Verlag. 8. Fan, W. Y., et al. 2001. Exp. Biol. Med. 226:1045. 9. Sitkauskiene, B., et al. 2005. Respir Res. 6:33.
IL-5 is a homodimeric, disulphide-linked protein produced by T-cells. Monomeric human IL-5 is a 126 amino acid protein with a reported molecular weight of 26 kD for the homodimeric protein. Mouse and human IL-5 are approximately 70% identical. IL-5 has been shown to promote the growth of immature hematopoietic BFU-E progenitors, stimulate the activation and differentiation of eosinophils, and promote the generation of cytotoxic lymphocytes. Mouse IL-5 induces the differentiation and proliferation of pre-activated B-cells and stimulates the production and secretion of IgM and IgA by B-cells stimulated with bacterial endotoxin. The TRFK5 antibody can neutralize the bioactivity of natural or recombinant IL-5.
Other Names:
Interleukin-5, Eosinophil differentiation factor (EDF), Eosinophil colony stimulating factor (Eo-CSF), B cell growth factor-2 (BCGF-2), B cell differentiation factor for IgM (BCDF-m), IgA enhancing factor (IgA-EF), T-cell replacing factor-1 (TRF-1)
Structure:
Cytokine; dimer; 40-50 kD (Mammalian)
Cellular Sources:
Mast cells, T cells, eosinophils
Cellular Targets:
Eosinophils, B cells
Receptors:
Heterodimer IL-5Rα (CD125); β-subunit (CDw131) in common with IL-3R, GM-CSFR
Bioactivity/Activities:
Growth/differentiation of B cells, eosinophils; proliferation/cell activation/differentiation of eosinophilic granulocytes
Antigen References:
1. Fitzgerald, K., et al. Eds. 2001. The Cytokine FactsBook. Academic Press, San Diego. 2. Takatsu, K., et al. 1988. Immunol. Rev. 102:107. 3. Takatsu, K. 1992. Curr. Opin. Immunol. 4:299. 4. Takatsu, K. 1991. Microbiol. Immunol. 35:593.
*These products may be covered by one or more Limited Use Label Licenses (see the BioLegend Catalog or our website, www.biolegend.com/terms). BioLegend products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without written approval of BioLegend. By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.
This data display is provided for general comparisons between formats. Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors. If you need assistance with selecting the best format contact our expert technical support team.
