TotalSeq™-B0945 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
Barcode Sequence
CCTATGAACGTAACG
Cat # Size Price Quantity Check Availability
502957 10 µg $369.00
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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 EDTA
Preparation
The antibody was purified by chromatography and conjugated with TotalSeq™-B oligomer 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

ICPG - Quality tested

Recommended Usage

Each lot of this antibody is quality control tested by intracellular immunofluorescent staining with flow cytometric analysis and the oligomer sequence is confirmed by sequencing. TotalSeq™- B antibodies are compatible with 10x Genomics Single Cell Gene Expression Solutions.

To maximize performance, it is strongly recommended that the reagent be titrated for each application, and that you centrifuge the antibody dilution before adding to the cells at 14,000xg at 2 - 8°C for 10 minutes. Carefully pipette out the liquid avoiding the bottom of the tube and add to the cell suspension. For Proteogenomics analysis, the suggested starting amount of this reagent for titration is ≤ 0.5 µg per million cells in 100 µL volume. Refer to the corresponding TotalSeq™ protocol for specific staining instructions. 


Buyer is solely responsible for determining whether Buyer has all intellectual property rights that are necessary for Buyer's intended uses of the BioLegend TotalSeq™ products. For example, for any technology platform Buyer uses with TotalSeq™, it is Buyer's sole responsibility to determine whether it has all necessary third party intellectual property rights to use that platform and TotalSeq™ with that platform.
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

Additional Product Notes

TotalSeq™ reagents are designed to profile protein levels at a single cell level following an optimized protocol similar to the CITE-seq workflow. A compatible single cell device (e.g. 10x Genomics Chromium System and Reagents) and sequencer (e.g. Illumina analyzers) are required. Please contact technical support for more information, or visit biolegend.com/totalseq.

The barcode flanking sequences are GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCTNNNNNNNNNN (PCR handle), and NNNNNNNNNGCTTTAAGGCCGGTCCTAGC*A*A (capture sequence). N represents either randomly selected A, C, G, or T, and * indicates a phosphorothioated bond, to prevent nuclease degradation.

View more applications data for this product in our Scientific Poster Library.

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)
RRID
AB_3068253 (BioLegend Cat. No. 502957)

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
Go To Top Version: 2    Revision Date: 03/18/2024

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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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