Ultra-LEAF™ Purified 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

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, ICFC - Quality tested
CyTOF®, ICC - Verified
IHC-F, WB, Neut - 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 2.0 - 6.0 µg/mL is recommended. To obtain a linear standard curve, serial dilutions of mouse TNF-α recombinant protein ranging from 500 to 4 pg/mL are recommended for each ELISA plate. It is recommended that the reagent be titrated for optimal performance for each application.

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

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. Qi S, et al. 2019. J Immunol. 203:3045. PubMed
2. McKnight Q, et al. 2020. J Bone Miner Res. 35:1352. PubMed
3. Carlson M, et al. 2017. Blood. 113(6):1365-74. PubMed
4. Kearney C, et al. 2017. Cell Death Differ.. 10.1038/cdd.2017.94. PubMed
5. Pham THM, et al. 2020. Cell Host & Microbe. 27(1):54-67.e5.. PubMed
6. Wood M, Rios D, Williams I 2016. Am J Physiol Cell Physiol. 311: C498 - C507. PubMed
7. Nie X et al. 2018. Neuron. 99(3):464-479 . PubMed
8. Iizasa E, et al. 2021. Nat Commun. 12:2299. PubMed
9. Jung H, et al. 2019. Autophagy. 15:1990. PubMed
10. Jain A, et al. 2020. Nat Immunol. 0.920138889. PubMed
11. Kurashima Y, et al. 2021. Nat Commun. 12:1067. PubMed

RRID

AB_11147367 (BioLegend Cat. No. 506331)
AB_11150777 (BioLegend Cat. No. 506332)
AB_2616671 (BioLegend Cat. No. 506347)
AB_2616672 (BioLegend Cat. No. 506348)

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

Documentation

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