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PerCP-Cy™5.5 Mouse Anti-Human TNF MAb11 RUO

PerCP-Cy™5.5 Mouse Anti-Human TNF 

Clone  MAb11   (RUO)

  • Brand BD Pharmingen™
  • Alternative Name Tumor necrosis factor alpha; TNF-a; TNF-α; TNFSF2; Cachectin
  • Vol. Per Test 5 µl
  • Isotype Mouse IgG1, κ
  • Reactivity Human (QC Testing)  Rhesus, Cynomolgus, Baboon (Tested in Development) 
  • Application Intracellular staining (flow cytometry) (Routinely Tested) 
  • Immunogen Recombinant Human TNF
  • Storage Buffer Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
  • Regulatory Status RUO
Product Details Recommended Assay References

Description

The MAb11 monoclonal antibody specifically binds to human tumor necrosis factor (TNF, also known as TNF-α) protein. TNF is an efficient juxtacrine, paracrine and endocrine mediator of inflammatory and immune functions. It regulates the growth and differentiation of a variety of cell types. TNF is cytotoxic for transformed cells when in conjunction with IFN-γ. It is secreted by activated monocytes/macrophages and other cells such as B cells, T cells and fibroblasts. The immunogen used to generate the MAb11 hybridoma was recombinant human TNF. The MAb11 antibody has been reported to crossreact with Rhesus Macaque TNF.

Format
  • Format PerCP-Cy™5.5
  • Excitation Source Blue 488 nm
  • Excitation Max 482 nm
  • Emission Max 678 nm

PerCP-Cy™5.5 is a tandem conjugate that combines PerCP with a cyanine dye. PerCP-Cy5.5 is not subject to photobeaching like PerCP and can be used with stream-in-air flow cytometers. It has less Fc receptor–mediated nonspecific staining than PE-Cy5. Additionally, the PerCP-Cy5.5 tandem conjugate is not as susceptible to fixative or light instability compared to APC-Cy7 and PE-Cy7.

Other Formats →

Suggested Companion Products

PerCP-Cy™5.5 Mouse IgG1 κ Isotype Control RUO
0.1 mg Cat No: 550795

BD Cytofix/Cytoperm Plus Kit (with BD GolgiStop) RUO
250 Tests Cat No: 554715

HiCK-1 Human Cytokine Positive Control Cells RUO
1 mL Cat No: 555061

BD Cytofix/Cytoperm™ Fixation/Permeablization Kit RUO
250 Tests Cat No: 554714

Protein Transport Inhibitor (Containing Monensin) RUO
0.7 mL Cat No: 554724

Recombinant Human TNF RUO
10 µg Cat No: 554618

Purified Mouse Anti-Human TNF MAb11 RUO
0.1 mg Cat No: 554510

Stain Buffer (FBS) RUO
500 mL Cat No: 554656

Stain Buffer (BSA) RUO
500 mL Cat No: 554657

Resources & Tools
 Spectrum Viewer  Panel Designer  Spectrum Viewer  Download TDS  Regulatory Document Website
Preparation and Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.

Product Notices

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  5. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  6. PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
  7. PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
  8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  9. Species testing during development may have been performed with a different format of the same clone. Selected applications have been tested for cross-reactivity.
  10. Cy is a trademark of GE Healthcare.
  11. Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.


Flow cytometry: The MAb11 antibody is useful for immunofluorescent staining and flow cytometric analysis to identify and enumerate TNF producing cells within mixed cell populations. A useful control investigators may consider using for demonstrating specificity of staining, is to pre-block with one of the following reagents: (1) recombinant human TNF (Cat. No. 554618) or (2) unlabeled MAb11 antibody (Cat. No. 554510), prior to staining.


  1. Black RA, Rauch CT, Kozlosky CJ, et al. A metalloproteinase disintegrin that releases tumour-necrosis factor-alpha from cells. Nature. 1997; 385(6618):729-733. View reference

  2. Danis VA, Franic GM, Rathjen DA, Brooks PM. Effects of granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-2, interferon-gamma (IFN-gamma), tumour necrosis factor-alpha (TNF-alpha) and IL-6 on the production of immunoreactive IL-1 and TNF-alpha by human monocytes. Clin Exp Immunol. 1991; 85(1):143-150. View reference

  3. Hogan MM, Vogel SN. Production of tumor necrosis factor by rIFN-gamma-primed C3H/HeJ (Lpsd) macrophages requires the presence of lipid A-associated proteins. J Immunol. 1988; 141(12):4196-4202. View reference

  4. Jaattela, M. . Biologic activities and mechanisms of action of tumor necrosis factor-α/cachectin. Lab Invest. 1991; 64:724-742.

  5. Jason J, Larned J. Single-cell cytokine profiles in normal humans: comparison of flow cytometric reagents and stimulation protocols. J Immunol Methods. 1997; 207(1):13-22. View reference

  6. Kriegler M, Perez C, DeFay K, Albert I, Lu SD. A novel form of TNF/cachectin is a cell surface cytotoxic transmembrane protein: ramifications for the complex physiology of TNF. Cell. 1988; 53(1):45-53. View reference

  7. Petyovka N, Lyach L, Voitenok NN. Homologous ELISA for detection of oligomeric human TNF: properties of the assay. J Immunol Methods. 1995; 186(2):161-170. View reference

  8. Prussin C, Metcalfe DD. Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies. J Immunol Methods. 1995; 188(1):117-128. View reference

  9. Rathjen DA, Cowan K, Furphy LJ, Aston R. Antigenic structure of human tumour necrosis factor: recognition of distinct regions of TNF alpha by different tumour cell receptors. Mol Immunol. 1991; 28(1-2):79-86. View reference

  10. Sander B, Andersson J, Andersson U. Assessment of cytokines by immunofluorescence and the paraformaldehyde-saponin procedure. Immunol Rev. 1991; 119:65-93. View reference

  11. Smith RA, Baglioni C. The active form of tumor necrosis factor is a trimer. J Biol Chem. 1987; 262(15):6951-6954. View reference

  12. Sopper S, Stahl-Hennig C, Demuth M, Johnston IC, Dorries R, ter Meulen V. Lymphocyte subsets and expression of differentiation markers in blood and lymphoid organs of rhesus monkeys. Cytometry. 1997; 29(4):351-362. View reference

  13. Verdier F, Aujoulat M, Condevaux F, Descotes J. Determination of lymphocyte subsets and cytokine levels in cynomolgus monkeys. Toxicology. 1995; 105(1):81-90. View reference

  14. Wang AM, Creasey AA, Ladner MB, et al. Molecular cloning of the complementary DNA for human tumor necrosis factor. Science. 1985; 228(4696):149-154. View reference

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