Biotin 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
HumanTNF-alpha_061604
  • HumanTNF-alpha_061604
  • Mab11_Biotin_TNFalpha_Antibody_ICC_011921
    Human PBMCs, stimulated with 1 µg/mL of LPS for 8 h and treated with Brefeldin A during the last 4 h, were prepared by cytospin, fixed and permeabilized on a slide and then treated with endogenous biotin blocking kit (Vector labs). Slides were stained with anti-human TNF-α biotin (clone MAb11) and DyLight™ 649 streptavidin (green) and counterstained with DAPI (blue). Images were acquired with an Olympus FV10i confocal microscope. Images courtesy of Teresa Rodriguez, Darius Schneider, and Matthias von Herrath, LaJolla Institute for Allergy and Immunology.
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502903 50 µg $101.00
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502904 500 µg $273.00
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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.
Preparation
The antibody was purified by affinity chromatography, and conjugated with biotin under optimal conditions.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C. Do not freeze.
Application

ELISA Detection - Quality tested
ICC - Verified
ELISPOT Detection, ICFC, FC - Reported in the literature, not verified in house

Recommended Usage

Each lot of this antibody is quality control tested by ELISA assay. For ELISA detection applications, a concentration range of 0.25 - 1.0 µg/mL is recommended. To obtain a linear standard curve, serial dilutions of TNF-α recombinant protein ranging from 500 to 4 pg/mL are recommended for each ELISA plate.

For use as an ELISPOT detection antibody, a concentration range of 0.5 - 2.0 µg/mL is recommended. For flow cytometric staining, the suggested use of this reagent is ≤ 0.25 µg per 106 cells in 100 µL volume. It is recommended that the reagent be titrated for optimal performance for each application.

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. Altemus J, et al. 2022. J Cell Physiol. 237:3001. PubMed
  2. Schwaderer J, et al. 2020. Cell Death Dis. 11:154. PubMed
  3. Choi C, et al. 2010. Anal Biochem. 405:1. PubMed
  4. Tao S, et al. 2020. Methods Mol Biol. 2108:181. PubMed
  5. Chan M, et al. 2021. Mol Syst Biol. 17:e10426. PubMed
RRID
AB_315255 (BioLegend Cat. No. 502903)
AB_315255 (BioLegend Cat. No. 502904)

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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