Purified anti-mouse IL-6 (Maxpar® Ready) Antibody

Product Details

Verified Reactivity

Mouse

Antibody Type

Monoclonal

Host Species

Rat

Immunogen

COS-7-expressed, recombinant mouse IL-6

Formulation

Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and EDTA.

Preparation

The antibody was purified by affinity chromatography.

Concentration

1.0 mg/ml

Storage & Handling

The antibody solution should be stored undiluted between 2°C and 8°C.

Application

ELISA Capture - Quality tested
CyTOF® - Verified

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. It is recommended that the reagent be titrated for optimal performance for each application.

This product is suitable for use with the Maxpar® Metal Labeling Kits. For metal labeling using Maxpar® Ready antibodies, proceed directly to the step to Partially Reduce the Antibody by adding 100 µl of Maxpar® Ready antibody to 100 µl of 4 mM TCEP-R in a 50 kDa filter and continue with the protocol. Always refer to the latest version of Maxpar® User Guide when conjugating Maxpar® Ready antibodies.

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. No. 504506).

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. No. 431301 to 431306) are specially developed and recommended.

Additional Product Notes

Maxpar® is a registered trademark of Standard BioTools Inc.

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

RRID

AB_2562845 (BioLegend Cat. No. 504509)

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

Can I obtain CyTOF data related to your Maxpar® Ready antibody clones?

We do not test our antibodies by mass cytometry or on a CyTOF machine in-house. The data displayed on our website is provided by Fluidigm^®. Please contact Fluidigm^® directly for additional data and further details.

http://techsupport.fluidigm.com/

Can I use Maxpar® Ready format clones for flow cytometry staining?

We have not tested the Maxpar^® Ready antibodies formulated in solution containing EDTA for flow cytometry staining. While it is likely that this will work in majority of the situations, it is best to use the non-EDTA formulated version of the same clone for flow cytometry testing. The presence of EDTA in some situations might negatively affect staining.

I am having difficulty observing a signal after conjugating a metal tag to your Maxpar® antibody. Please help troubleshoot.

We only supply the antibody and not test that in house. Please contact Fluidigm^® directly for troubleshooting advice: http://techsupport.fluidigm.com/

Is there a difference between buffer formulations related to Maxpar® Ready and purified format antibodies?

The Maxpar^® Ready format antibody clones are formulated in Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and EDTA. The regular purified format clones are formulated in solution that does not contain any EDTA. Both formulations are however without any extra carrier proteins.

Other Formats

Certificate of Analysis