Brilliant Violet 605™ anti-mouse TNF-α Antibody

Product Details

Verified Reactivity

Mouse

Antibody Type

Monoclonal

Host Species

Rat

Immunogen

E. coli-expressed, recombinant mouse TNF-α

Formulation

Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and BSA (origin USA).

Preparation

The antibody was purified by affinity chromatography and conjugated with Brilliant Violet 605™ under optimal conditions.

Concentration

Lot-specific (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, and protected from prolonged exposure to light. Do not freeze.

Application

ICFC - Quality tested

Recommended Usage

Each lot of this antibody is quality control tested by intracellular immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is ≤0.5 µg per million cells. It is recommended that the reagent be titrated for optimal performance for each application.

Brilliant Violet 605™ excites at 405 nm and emits at 603 nm. The bandpass filter 610/20 nm is recommended for detection, although filter optimization may be required depending on other fluorophores used. Be sure to verify that your cytometer configuration and software setup are appropriate for detecting this channel. Refer to your instrument manual or manufacturer for support. Brilliant Violet 605™ is a trademark of Sirigen Group Ltd.

Learn more about Brilliant Violet™.

This product is subject to proprietary rights of Sirigen Inc. and is made and sold under license from Sirigen Inc. The purchase of this product conveys to the buyer a non-transferable right to use the purchased product for research purposes only. This product may not be resold or incorporated in any manner into another product for resale. Any use for therapeutics or diagnostics is strictly prohibited. This product is covered by U.S. Patent(s), pending patent applications and foreign equivalents.

Excitation Laser

Violet Laser (405 nm)

Application Notes

ELISA or ELISPOT Detection: The biotinylated MP6-XT22 antibody is useful as a detection antibody for a sandwich ELISA or ELISPOT assay, when used in conjunction with purified 6B8 antibody (Cat. Nos. 510802 & 510804) as the capture antibody.
ELISA Capture: The purified MP6-XT22 antibody is useful as the capture antibody in a sandwich ELISA when used in conjunction with the biotinylated Poly5160 antibody (Cat. No. 516003) as the detection antibody and recombinant mouse TNF-α (Cat. No. 575209) as the standard.
Flow Cytometry^6,11,12:The fluorochrome-labeled MP6-XT22 antibody is useful for intracellular immunofluorescent staining and flow cytometric analysis to identify TNF-a-producing cells within mixed cell populations.
Neutralization^1,5,10,16,17:The MP6-XT22 antibody can neutralize the bioactivity of natural or recombinant TNF-α. The LEAF™ purified antibody (Endotoxin < 0.1 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for neutralization of mouse TNF-α bioactivity in vivo and in vitro(Cat. No. 506310). For in vivo studies or highly sensitive assays, we recommend Ultra-LEAF™ purified antibody (Cat. No. 506332) with a lower endotoxin limit than standard LEAF™ purified antibodies (Endotoxin < 0.01 EU/µg).
Additional reported applications (for the relevant formats) include: Western blotting, immunohistochemical staining of paraformaldehyde-fixed, saponin-treated frozen tissue sections^7-9 in vivo detection⁵, immunofluorescence, and immunocytochemistry.
Note: For testing mouse TNF-α in serum, plasma or supernatant, BioLegend's ELISA Max™ Sets (Cat. No. 430901) are specially developed and recommended.

Application References

(PubMed link indicates BioLegend citation)

1. Abrams J, et al. 1992. Immunol. Rev. 127:5. (Neut)
2. Abrams J, et al. 1995. Curr. Prot. Immunol. John Wiley and Sons, New York. Unit 6.20
3. Mo X, et al. 1995. J. Virol. 69:1288.
4. Sarawar S, et al. 1994. J. Immunol. 153:1246.
5. Via C, et al. 2001. J. Immunol. 167:6821. (Neut)
6. Infante-Duarte C, et al. 2000 J. Immunol. 165:6107. (FC)
7. Jacobs M, et al. 2000. Immunology 100:494. (IHC)
8. Marinova-Mutachieva L, et al. 1997. Clin. Exp. Immunol. 107:507. (IHC)
9. Williams RO, et al. 2000. J. Immunol. 165:7240. (IHC)
10. Scanga CA, et al. 1999. Infect. Immun. 67:4531. (Neut)
11. Akilov OE, et al. 2007. J. Leukoc. Biol. 2007;10.1189/jlb.0706439. (FC)
12. Lawson BR, et al. 2007. J. Immunol. 178:5366. (FC)
13. Patole PS, et al. 2005. J. Am. Soc. Nephrol. 16:3273. PubMed
14. Wu S, et al. 2005. Neurosci Lett. 394:158. PubMed
15. Carlson MJ, et al. 2009. Blood 113:1365. PubMed
16. Shivakumar P, et al. 2017. JCI Insight. 2:e88747 1. PubMed
17. Kearney CJ, et al. 2017. Cell Death Differ. 10.1038/cdd.2017.94. PubMed

Product Citations

1. Bruno MEC, et al. 2022. J Cell Physiol. 237:706. PubMed
2. Grebinoski S, et al. 2022. Nat Immunol. 23:868. PubMed
3. Hao J, et al. 2022. Cell Rep. 41:111804. PubMed
4. Wu Z, et al. 2023. EMBO Rep. 24:e56524. PubMed
5. Le DT, et al. 2023. iScience. 26:106059. PubMed
6. Tursi NJ, et al. 2023. Front Immunol. 14:1072810. PubMed
7. Verma S, et al. 2016. J Virol. 90: 650 - 658. PubMed
8. Konrath KM, et al. 2022. Cell Rep. 38:110318. PubMed
9. Hassan AO, et al. 2019. Cell Rep. 28:2634. PubMed
10. Dikiy S, et al. 2021. Immunity. 54(5):931-946.e11. PubMed
11. He X, et al. 2022. Cancer Immunol Res. 10:314. PubMed
12. Xu Z, et al. 2020. Cancer Immunol Res. 1354:8. PubMed
13. Campbell C et al. 2018. Immunity. 48(6):1245-1257 . PubMed
14. Hassan AO, et al. 2020. Cell. 183:169. PubMed
15. Bachy S, et al. 2022. iScience. 25:103758. PubMed
16. Maulloo CD, et al. 2021. Front Immunol. 12:714842. PubMed
17. Evgin L, et al. 2022. Sci Transl Med. 14:eabn2231. PubMed
18. Lai Y, et al. 2022. Clin Transl Med. 12:e999. PubMed
19. He X, et al. 2021. J Immunother Cancer. 9:. PubMed
20. Loo Yau H, et al. 2021. Molecular Cell. 81(7):1469-1483.e8. PubMed
21. Ying B, et al. 2022. Sci Transl Med. 14:eabm3302. PubMed
22. Conforti A, et al. 2021. Mol Ther. S1525-0016:00466. PubMed
23. Laczkó D, et al. 2020. Immunity. 53:724. PubMed

RRID

AB_11123912 (BioLegend Cat. No. 506329)

Antigen Details

Structure

TNF superfamily; dimer/trimer; 17.5-150 kD (Mammalian)

Bioactivity

Paracrine/endocrine mediator of inflammatory and immune functions; selectively cytotoxic for transformed cells; endothelial cell alterations; chemoattractant

Cell Sources

Activated monocytes, neutrophils, macrophages, T cells, B cells, NK cells, LAK cells

Cell Targets

Monocytes, neutrophils, macrophages, T cells, fibroblasts, endothelial cells, osteoclasts, adipocytes, astroglia, microglia

Receptors

TNFRSF1A (TNF-R1, CD120a, TNFR-p60 Type β, p55); TNFRSF1B (TNF-R2, CD120b, TNFR-p80 Type A, p75)

Cell Type

Tregs

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. Beutler B, et al. 1988. Annu. Rev. Biochem. 57:505.
3. Beutler B, et al. 1989. Annu. Rev. Immunol. 7:625.
4. Tracey K, et al. 1993. Crit. Care Med. 21:S415.

Regulation

Processed by TACE for secretion; upregulated by interferons, IL-2, GM-CSF, substance P, bradykinin, PAF, immune complexes, and cyclooxygenase; downregulated by IL-6, TGF-β, vitamin D3, prostaglandin E2, and PAF antagonists

Gene ID

21926 View all products for this Gene ID

UniProt

View information about TNF-alpha on UniProt.org

Related Protocols

• Surface and Intracellular Cytokine Staining for Flow Cytometry - Video
• Intracellular Flow Cytometry Staining Protocol

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