mouse/human IL-5 antibody, Functional Grade Purified conjugate

Contents: Functional Grade Purified anti-mouse/human interleukin-5 (IL-5)
Catalog Number: 16-7052
Formulation: Phosphate buffer pH 7.2,
150 mM NaCl, No NaN₃
Storage Conditions: Store at 4°C.
Avoid repeated freeze/thaw cycles.
KEEP CONTENTS STERILE.
Endotoxin Level: Less than 0.001 ng/ug antibody, as determined by the LAL assay.
Clone: TRFK5
Isotype: Rat IgG1, κ

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16-7052-81

50 ug

16-7052-85

500 ug

Naive mouse splenocytes (SJL/J strain) were stimulated with 2 ug/ml ConA in HL-1 media for 48 hours (right well) in a mouse IL-5 ELISPOT assay plate (pre-coated with functional grade TRFK5 antibody). Left well is medium alone control.

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
14-7052	Affinity Purified anti-mouse/human IL-5 (Interleukin-5, IL5)	N/A	N/A	ELISA cap ELISPOT cap IC Flow NU
16-7052	Functional Grade* Purified anti-mouse/human IL-5 (Interleukin-5, IL5)	N/A	N/A	NU
12-7052	Phycoerythrin (PE) anti-mouse/human IL-5 (Interleukin-5, IL5)	488	575	IC Flow
*Functional Grade™ (FG™) Purified: Azide-free, sterile-filtered, and endotoxin < 0.001 ng/µg (unless otherwise noted). *Functional Grade™ (FG™) Biotin: Azide-free, sterile-filtered, and endotoxin < 0.05 ng/µg. Purified: Contains azide, not sterile-filtered, and not endotoxin tested.
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Pacific Blue® and APC-Alexa Fluor® 750 conjugated products with eFluor™ 450 and APC-eFluor™ 780 conjugated products, respectively.

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Description

The TRFK5 antibody reacts with mouse and human interleukin-5 (IL-5). The TRFK5 antibody is a neutralizing antibody. Mouse IL-5 is a disulfide-linked homodimer, containing two 113 amino acid peptides; these resolve as 32-34 kDa bands in SDS-PAGE. IL-5 is produced by T cells and has been known as eosinophil-differentiating factor (EDF), B cell growth factor II (BCGFII), and T cell-replacing factor (TRF). IL-5 induces eosinophil differentiation and promotes eosinophil survival and activation. In mice, IL-5 has been shown to stimulate B cell proliferation and antibody production.

Applications Reported

For research use only, not for diagnostic or therapeutic use. The TRFK5 antibody has been reported for use in ELISA capture, ELISPOT capture, neutralization of IL-5 bioactivity, IHC, and intracellular staining for flow cytometric analysis.

Applications Tested

The Functional Grade Purified TRFK5 antibody has been tested by LAL assay to verify low endotoxin levels, by ELISA, and in bioassay for neutralization of mouse IL-5 bioactivity.

The TRFK5 antibody at 0.06 ug/ml has been found to inhibit by 50% the biological effects of 1.0 ng/ml mouse IL-5 in an MC/9 cell proliferation assay. Detailed information and protocols about cytokine bioassays and in vitro cytokine neutralization using antibodies can be found in the BestProtocols section.

The TRFK5 antibody has been tested as the capture antibody in a sandwich ELISA for analysis of mouse IL-5 in combination with the biotin TRFK4 (13-7051) antibody for detection and recombinant mouse IL-5 (14-8051) as the standard. A suitable range of concentrations of this antibody for ELISA capture is 1-4 µg/ml. A standard curve consisting of doubling dilutions of the recombinant standard over the range of 1000 pg/ml - 8 pg/ml should be included in each ELISA plate.

The TRFK5 antibody has been tested as the capture antibody in a sandwich ELISA for analysis of human IL-5 in combination with the biotinylated JES1-5A10 (13-7059) antibody for detection and recombinant human IL-5 (\lt{14-80591}) as the standard. A suitable range of concentrations of this antibody for ELISA capture is 1-4 µg/ml. A standard curve consisting of doubling dilutions of the recombinant standard over the range of 1000 pg/ml - 8 pg/ml should be included in each ELISA plate.

References

Abrams, J. S., M. G. Roncarolo, et al. (1992). "Strategies of anti-cytokine monoclonal antibody development: immunoassay of IL-10 and IL-5 in clinical samples." Immunol Rev 127: 5-24.
Schumacher, J. H., A. O'Garra, et al. (1988). "The characterization of four monoclonal antibodies specific for mouse IL-5 and development of mouse and human IL-5 enzyme-linked immunosorbent." J Immunol141(5): 1576-81.

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