Alexa Fluor® 647 anti-human TNF-α Antibody

Pricing & Availability
Clone
MAb11 (See other available formats)
Regulatory Status
RUO
Other Names
Tumor necrosis factor-α, Cachectin, Necrosin, Macrophage cytotoxic factor (MCF), Differentiation inducing factor (DIF), TNFSF2
Isotype
Mouse IgG1, κ
A_Mab11_A647_070609.jpg
PMA/ionomycin-stimulated (6 hours) human peripheral blood lymphocytes stained with mouse IgG1 Alexa Fluor® 647 isotype control and CD3 (UCHT1) PE
  • A_Mab11_A647_070609.jpg
    PMA/ionomycin-stimulated (6 hours) human peripheral blood lymphocytes stained with mouse IgG1 Alexa Fluor® 647 isotype control and CD3 (UCHT1) PE
  • B_Mab11_A647_070609.jpg
    PMA/ionomycin-stimulated (6 hours) human peripheral blood lymphocytes stained with MAb11 Alexa Fluor® 647 and CD3 (UCHT1) PE
Compare all formats See Alexa Fluor® 647 spectral data
Cat # Size Price Quantity Check Availability
502916 100 tests $265.00
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Description

TNF-α is secreted by macrophages, monocytes, neutrophils, T cells, and NK cells. Many transformed cell lines also secrete TNF-α. Monomeric human TNF-α is a 157 amino acid protein (non-glycosylated) with a reported molecular weight of 17 kD. TNF-α forms multimeric complexes; stable trimers are most common in solution. A 26 kD membrane form of TNF-α has also been described. TNF-α binding to surface receptors elicits a wide array of biological activities including: cytolysis and cytostasis of many tumor cell lines in vitro, hemorraghic necrosis of tumors in vivo, increased fibroblast proliferation, and enhanced chemotaxis and phagocytosis in neutrophils.

Technical data sheet

Product Details

Verified Reactivity
Human
Reported Reactivity
Cat, Chimpanzee, Baboon, Cynomolgus, Rhesus, Pigtailed Macaque, Sooty Mangabey, Pig
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
E. coli-expressed, recombinant human 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 Alexa Fluor® 647 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 5 µL per 106 cells in 100 µL volume. It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 647 has a maximum emission of 668 nm when it is excited at 633nm / 635nm.


Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.

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Excitation Laser
Red Laser (633 nm)
Application Notes

ELISA or ELISPOT Detection: The biotinylated MAb11 antibody is useful as the detection antibody in a sandwich ELISA or ELISPOT, when used in conjunction with the purified MAb1 antibody (Cat. No. 502802/502804) as the capture antibody.

Flow Cytometry3,5,6,10: The fluorochrome-labeled MAb11 antibody is useful for intracellular and membrane-bound immunofluorescent staining and flow cytometric analysis to identify TNF-a-producing cells within mixed cell populations.

Additional reported applications (for the relevant formats) include: neutralization1,2, immunohistochemical staining of paraformaldehyde-fixed, saponin-treated frozen tissue sections4 and acetone-fixed frozen tissue sections8, immunocytochemistry7, and immunofluorescence9. The MAb11 antibody can neutralize the bioactivity of natural or recombinant TNF-a.

Note: For testing human TNF-a in serum or plasma, BioLegend's ELISA Max™ Sets (Cat. No. 430201 to 430206) are specially developed and recommended. The LEAF™ purified antibody (Endotoxin <0.1 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for neutralization of human TNF-a bioactivity (Cat. No. 502922).

The Purified MAb1 antibody is useful in neutralization2 and as the capture antibody in a sandwich ELISA or ELISPOT assay, when used in conjunction with the biotinylated MAb11 antibody (Cat. No. 502904/502914) as the detecting antibody.

Clone MAb11 cross-reacts to Cat11

Application References

(PubMed link indicates BioLegend citation)
  1. Rathjen D, et al. 1991. Mol. Immunol. 28:79. (Neut)
  2. Ablamunits V, et al. 2010. Eur. J. Immunol. 40:2891. (Neut)
  3. Enr quez J, et al. 2002. Adv. Perit. Dial. 18:177. (ICFC)
  4. Andersson U, et al. 1999. Detection and quantification of gene expression. New York:Springer-Verlag. (IHC)
  5. Chen H, et al. 2005. J. Immunol. 175:591. (ICFC)
  6. Iwamoto S, et al. 2007. J. Immunol. 179:1449. (ICFC) PubMed
  7. Andersson U, et al. 2000. J. Exp. Med. 192:565. (ICC)
  8. Moormann AM, et al. 1999. J. Infect. Dis. 180:1987. (IHC)
  9. Zhao XJ, et al. 2003. J. Immunol. 170:2923. (IF)
  10. Rieger R, et al. 2009. Cancer Gene Ther. 1:53-64. (FC)
  11. Maksaereekul S, et al. 2009. Vaccine. 28:3754 (FC)
Product Citations
  1. Porbahaie M, et al. 2023. PLoS One. 18:e0279626. PubMed
  2. Franzoni G, et al. 2013. Clin Vaccine Immunol. 20:1604. PubMed
  3. Fang F, et al. 2022. JCI Insight. 7:. PubMed
  4. Muenchhoff M, et al. 2016. Sci Transl Med. 8: 358ra125. PubMed
  5. Lee J, et al. 2020. Sci Rep. 10:17753. PubMed
  6. Jacquelot N, et al. 2016. J Invest Dermatol. 136: 994-1001. PubMed
  7. Vacaflores A, et al. 2016. PLoS One. 11: 0157175. PubMed
  8. Huang RS, et al. 2021. Curr Protoc. 1:e246. PubMed
  9. Fang F, et al. 2021. Cell Rep. 37:109981. PubMed
  10. Ricklin ME, et al. 2017. Sci Rep. 7:16379. PubMed
  11. Yin S, et al. 2015. Sci Rep. 5: 14432. PubMed
  12. Auray G, et al. 2016. J Immunol. 197(12):4791-4806. PubMed
  13. Harris LD, et al. 2020. Front Cell Infect Microbiol. 10:120. PubMed
RRID
AB_493123 (BioLegend Cat. No. 502916)

Antigen Details

Structure
TNF superfamily; dimer/trimer; 17 kD (Mammalian)
Bioactivity
Paracrine/endocrine mediator of inflammatory and immune functions; selectively cytotoxic for transformed cells; 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
Neutrophils, Tregs
Biology Area
Cell Biology, Immunology, Innate Immunity, Neuroinflammation, Neuroscience
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
Type II integral membrane protein processed by TACE for secretion; upregulated by interferons, IL-2, GM-CSF, substance P, bradykinin, PAF, immune complexes, cyclooxygenase; downregulated by IL-6, TGF-β, vitamin D3, prostaglandin E2, PAF antagonists
Gene ID
7124 View all products for this Gene ID
UniProt
View information about TNF-alpha on UniProt.org
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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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