Ultra-LEAF™ Purified anti-mouse IL-10 Antibody

Product Details

Verified Reactivity

Mouse

Antibody Type

Monoclonal

Host Species

Rat

Immunogen

E. coli-expressed, recombinant mouse IL-10

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
ELISPOT Capture, Neut, 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 and ELISPOT capture applications, a concentration range of 4-8 µg/ml is recommended. To obtain a linear standard curve, serial dilutions of IL-10 recombinant protein ranging from 2000 to 15 pg/ml are recommended for each ELISA plate. It is recommended that the reagent be titrated for optimal performance for each application.

* For ELISA/ELISPOT capture, it is very critical to use 0.2 M Sodium Phosphate Buffer, pH 6.5 (11.8g Na₂HPO₄, 16.1g NaH₂PO₄; q.s. to 1.0 L) as coating buffer.

Note: Carbonate buffer, pH 9.5 should not be used as coating buffer for JES5-2A5. It may cause high background and lower sensitivity.

Application Notes

ELISA or ELISPOT Capture^1-5,7,10: The purified JES5-2A5 antibody is useful as the capture antibody in a sandwich ELISA or ELISPOT assay, when used in conjunction with the biotinylated JES5-16E3 antibody (Cat. No. 505004) as the detecting antibody. The LEAF™ purified antibody is suggested for ELISPOT capture.

ELISA or ELISPOT Detection: The biotinylated JES5-2A5 antibody is useful as the detecting antibody in a sandwich ELISA or ELISPOT assay, when used in conjunction with the Purified JES5-16E3 antibody (Cat. No. 505002/505012) as the capture antibody.

Neutralization^2,8,9: The LEAF™ purified antibody (Endotoxin <0.1 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for neutralization of mouse IL-10 bioactivity in vivo and in vitro (Cat. No. 504904).

Additional reported applications (for the relevant formats) include: Western blotting¹² and in vivo capture⁶.

Note: For testing mouse IL-10 in serum, plasma or supernatant, BioLegend's ELISA Max™ Sets (Cat. No. 431401 to 431406) are specially developed and recommended.

Application References

(PubMed link indicates BioLegend citation)

1. Sander B, et al. 1993. J. Immunol. Meth. 166:201.
2. Abrams J, et al. 1992. Immunol. Rev. 127:5.
3. Abrams J, 1995. Curr. Prot. Immunol. John Wiley and Sons, New York. Unit 6.20.
4. Mo X, et al. 1995. J. Virol. 69:1288.
5. Sarawar S, et al. 1994. J. Immunol. 153:1246.
6. Finkelman F, et al. 2003. Curr. Prot. Immunol. John Wiley & Sons, New York. Unit 6.28.
7. Brummel R and Lenert P, 2005. J. Immunol. 174:2429.
8. Hara M, et al. 2001. J. Immunol. 166:3789.
9. Riemann M, et al. 2005. J. Immunol. 175:3560.
10. Xu G, et al. 2007. J. Immunol. 179:5358.
11. Guo Z, et al. 2013. J. Immunol. 190:4337. PubMed
12. van der Poll T, et al. 1995. J. Immunol. 155:5397. (WB)

Product Citations

1. Song J, et al. 2022. Clin Transl Med. 12:e992. PubMed
2. Shute A, et al. 2021. Microbiome. 9:186. PubMed

RRID

AB_2810629 (BioLegend Cat. No. 504912)
AB_2810629 (BioLegend Cat. No. 504910)
AB_2810629 (BioLegend Cat. No. 504911)
AB_2810629 (BioLegend Cat. No. 504907)
AB_2810629 (BioLegend Cat. No. 504908)
AB_2810629 (BioLegend Cat. No. 504909)

Antigen Details

Structure

Acid-labile cytokine; dimer; 17-21 kD (Mammalian)

Bioactivity

Inhibits IFN-γ, TNF-β, IL-2 production by T_H1 clones; inhibits macrophage-mediated IL-1, IL-6, TNF-α synthesis; suppresses delayed type hypersensitivity response; stimulates T_H2 cell response; mast cell proliferation

Cell Sources

Activated CD8⁺ T cells, T_H0, T_H2 subset of CD4⁺ T cells, Ly-1⁺ B cells, monocytes, macrophages, keratinocytes

Cell Targets

T cells, B cells, mast cells, macrophages

Receptors

IL-10R (CDw210)

Cell Type

Tregs

Biology Area

Immunology

Molecular Family

Cytokines/Chemokines

Antigen References

1. Fitzgerald, K., et al. Eds. 2001. The Cytokine FactsBook. Academic Press, San Diego.
2. de Waal-Malefy, T, et al. 1992. Curr. Opin. Immunol. 4:314.
3. Howard, M., et al. 1992. Immunol. Today. 13:198.
4. Quesniaux, V. 1992. Research Immunol. 143:385.

Regulation

Downregulated by IL-4, IL-10

Gene ID

16153 View all products for this Gene ID

UniProt

View information about IL-10 on UniProt.org

Documentation

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

Reviews

Related Protocols

• 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