Ultra-LEAF™ Purified anti-mouse IL-6 Antibody

Product Details

Verified Reactivity

Mouse

Antibody Type

Monoclonal

Host Species

Rat

Immunogen

COS-7-expressed, recombinant mouse IL-6

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, IHC-F - Reported in the literature, not verified in house

Recommended Usage

Each lot of this antibody is quality control tested by ELISA assay. For ELISA applications, a concentration range of 0.5 – 2.0 µg/mL is recommended. To obtain a linear standard curve, serial dilutions of mouse IL-6 recombinant protein ranging from 500 to 8 pg/mL are recommended for each ELISA plate. For ELISPOT applications, a concentration range of 4.0 - 8.0 µg/mL is recommended. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

ELISA Capture^1-3,11 or ELISPOT Capture^4,5: The purified MP5-20F3 antibody is useful as the capture antibody in a sandwich ELISA or ELISPOT assay, when used in conjunction with the biotinylated MP5-32C11 antibody (Cat. No. 504602) as the detecting antibody and recombinant mouse IL-6 (Cat. No. 575709) as the standard. The Ultra-LEAF™ purified antibody is suggested for ELISPOT capture.

Flow Cytometry: The fluorochrome-labeled MP5-20F3 antibody is useful for intracellular immunofluorescent staining and flow cytometric analysis to identify IL-6 -producing cells within mixed cell populations. For intracellular cytokine staining protocol, please visit www.biolegend.com and click on the support section.

Neutralization^1-2,8-10: The Ultra-LEAF™ purified antibody (Endotoxin <0.1 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for neutralization of mouse IL-6 bioactivity in vivo and in vitro (Cat. Nos. 504511-504516).

Additional reported applications (for relevant formats of this clone) include: In Vivo Capture⁷, and immunohistochemical staining (paraformaldehyde-fixed/saponin-treated⁶, and acetone-fixed frozen tissue sections^{13, 14}).

Note: For testing mouse IL-6 in serum, plasma or supernatant, BioLegend's ELISA Max™ Sets (Cat. Nos. 431301 to 431306) are specially developed and recommended.

Application References

(PubMed link indicates BioLegend citation)

1. Abrams J, et al. 1992. Immunol. Rev. 127:5. (ELISA Capture, Neut)
2. Sander B, et al. 1993. J. Immunol. Meth. 166:201. (ELISA Capture, Neut)
3. Abrams J, et al. 1995. Curr. Prot. Immunol. John Wiley and Sons, New York. Unit 6.20. (ELISA Capture)
4. Fujihashi K, et al. 1991. J. Immunol. Meth. 160:181. (ELISPOT Capture)
5. Klinman D, et al. 1994. Curr. Prot. Immunol. John Wiley and Sons, New York. Unit 6.19. (ELISPOT Capture)
6. Litton M, et al. 1994. J. Immunol Meth. 175:47. (IHC)
7. Finkelman F, et al. 2003. Curr. Prot. Immunol. John Wiley & Sons, New York. Unit 6.28. (In Vivo Capture)
8. Starnes HF Jr, et al. 1990. J. Immunol. 145:4185. (Neut)
9. Riedemann NC, et al. 2003. J. Immunol. 170:503. (Neut)
10. Rochman I, et al. 2005. J. Immunol. 174:4761. (Neut)
11. O'Connell PJ, et al. 2006. Blood 107:1010. (ELISA Capture)
12. Ozeki Y, et al. 2010. Int. Immunol. 22:179. PubMed
13. Galie M, et al. 2005. Carcinogenesis 26:1868 (IHC)
14. Schone MP, et al. 2000. J Immunol 165:6583 (IHC)
15. Keswani T, et al. 2014. Exp Parasitol. 141:82. PubMed
16. Notley CA, et al. 2015. J Immunol. 194:1621. PubMed

Product Citations

1. Cui L, et al. 2022. J Biomed Sci. 29:101. PubMed
2. Yang L, et al. 2023. Sci Rep. 13:1827. PubMed
3. Zioni N, et al. 2023. Nat Commun. 14:2070. PubMed
4. Kuczynski EA, et al. 2023. STAR Protoc. 102144:4. PubMed
5. Izumi G, et al. 2021. Nat Commun. 12:5029. PubMed

RRID

AB_2814417 (BioLegend Cat. No. 504516)
AB_2814417 (BioLegend Cat. No. 504514)
AB_2814417 (BioLegend Cat. No. 504515)
AB_2814417 (BioLegend Cat. No. 504511)
AB_2814417 (BioLegend Cat. No. 504512)
AB_2814417 (BioLegend Cat. No. 504513)

Antigen Details

Structure

Neuropoietic cytokine family; 22-29 kD (Mammalian)

Bioactivity

Hematopoiesis; antigen-specific immune responses, inflammatory reactions; acute phase reaction

Cell Sources

T cells, B cells, macrophages, bone marrow stromal cells, fibroblasts, keratinocytes, mesangium, astrocytes, endothelial cells

Cell Targets

T cells, B cells, hepatocytes, cholinergic neurons

Receptors

Heterodimer IL-6Rα (CD126)/IL-6Rβ (CD130; gp130)

Biology Area

Immunology, Innate Immunity

Molecular Family

Cytokines/Chemokines

Antigen References

1. Fitzgerald K, et al. Eds. 2001. The Cytokine FactsBook. Academic Press, San Diego.
2. Hirano T. 1998. Intl. Rev. Immunol. 16:249.
3. Patterson P. 1992. Curr. Opin. Neurobiol. 2:94.
4. Van Oers M, et al. 1993. Ann. Hematology 66:219.

Regulation

Upregulated by IL-1, TNF, PDGF, IFN-β, TNF-α, NGF, IL-17; downregulated by glucocorticoids IL-4, TGF-β

Gene ID

16193 View all products for this Gene ID

UniProt

View information about IL-6 on UniProt.org

Documentation

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

Reviews

Related Protocols

• Active Protocols: Sandwich ELISA - Video
• Sandwich ELISA 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