Ultra-LEAF™ Purified anti-mouse/human IL-5 Antibody

Product Details

Verified Reactivity

Mouse, Human

Reported Reactivity

Guinea Pig

Antibody Type

Monoclonal

Host Species

Rat

Immunogen

Partially-purified T cell clone supernatant

Formulation

0.2 µm filtered in phosphate-buffered solution, pH 7.2, containing no preservative.

Endotoxin Level

Less than 0.01 EU/µg of the protein (< 0.001 ng/µg of the protein) as determined by the LAL test.

Preparation

The Ultra-LEAF™ (Low Endotoxin, Azide-Free) antibody was purified by affinity chromatography.

Concentration

The antibody is bottled at the concentration indicated on the vial, typically between 2 mg/mL and 3 mg/mL. Older lots may have also been bottled at 1 mg/mL. To obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.

Storage & Handling

The antibody solution should be stored undiluted between 2°C and 8°C. This Ultra-LEAF™ solution contains no preservative; handle under aseptic conditions.

Application

ELISA Capture - Quality tested
CyTOF® - Verified
ELISPOT Capture, Neut, ICFC, IHC, WB - Reported in the literature, not verified in house

Recommended Usage

Each lot of this antibody is quality control tested by ELISA assay. For ELISA capture applications, a concentration range of 1.0 - 4.0 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. No. 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. No. 501002). It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

ELISA^1,3,4 or ELISPOT^2,6 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) as the detection antibody for human IL-5. The Ultra-LEAF™ purified antibody is suggested for ELISPOT capture.

Flow Cytometry: The fluorochrome-labeled TRFK51 antibody is useful for intracellular immunofluorescent staining and flow cytometric analysis to identify IL-5-producing cells within mixed cell populations.

Neutralization^1,9: The Ultra-LEAF™ purified antibody (Endotoxin <0.1 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for neutralization of human IL-5 bioactivity (Cat. Nos. 504313-504318).

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 and 430404) are specially developed and recommended. For testing mouse IL-5 in serum, plasma or supernatant, BioLegend's ELISA MAX™ Sets (Cat. Nos. 431201 and 431206) are specially developed and recommended.

Application References

(PubMed link indicates BioLegend citation)

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.

RRID

AB_2814411 (BioLegend Cat. No. 504316)
AB_2814411 (BioLegend Cat. No. 504317)
AB_2814411 (BioLegend Cat. No. 504318)
AB_2814411 (BioLegend Cat. No. 504313)
AB_2814411 (BioLegend Cat. No. 504314)
AB_2814411 (BioLegend Cat. No. 504315)

Antigen Details

Structure

Cytokine; dimer; 40-50 kD (Mammalian)

Bioactivity

Growth/differentiation of B cells, eosinophils; proliferation/cell activation/differentiation of eosinophilic granulocytes

Cell Sources

Mast cells, T cells, eosinophils

Cell Targets

Eosinophils, B cells

Receptors

Heterodimer IL-5Rα (CD125); β-subunit (CDw131) in common with IL-3R, GM-CSFR

Biology Area

Immunology

Molecular Family

Cytokines/Chemokines

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.

Gene ID

16191 View all products for this Gene ID 3567 View all products for this Gene ID

UniProt

View information about IL-5 on UniProt.org

Documentation

• Technical Data Sheet (pdf)
• Certificate of Analysis
• Safety Data Sheet

Reviews

Related Protocols

• Cell Surface Flow Cytometry Staining Protocol

Related Pages & Pathways

Related FAQs

Do you guarantee that your antibodies are totally pathogen free?

BioLegend does not test for pathogens in-house aside from the GoInVivo™ product line. However, upon request, this can be tested on a custom basis with an outside, independent laboratory.

Does BioLegend test each Ultra-LEAF™ antibody by functional assay?

No, BioLegend does not test Ultra-LEAF™ antibodies by functional assays unless otherwise indicated. Due to the possible complexities and variations of uses of biofunctional antibodies in different assays and because of the large product portfolio, BioLegend does not currently perform functional assays as a routine QC for the antibodies. However, we do provide references in which the antibodies were used for functional assays and we do perform QC to verify the specificity and quality of the antibody based on our strict specification criteria.

Does BioLegend test each Ultra-LEAF™ antibody for potential pathogens?

No, BioLegend does not test for pathogens in-house unless otherwise indicated.  However, we can recommend an outside vendor to perform this testing as needed.

Have you tested this Ultra-LEAF™ antibody for in vivo or in vitro applications?

We don't test our antibodies for in vivo or in vitro applications unless otherwise indicated. Depending on the product, the TDS may describe literature supporting usage of a particular product for bioassay. It may be best to further consult the literature to find clone specific information.

Other Formats

Certificate of Analysis