Clone:
REA656
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
ICFC
Alternative names:
Tumor necrosis factor, Cachectin, TNF-alpha, TNF-a, DIF-alpha, TNFa, TNFSF2

Extended validation for TNF-α Antibody, anti-human, REAfinity™

Specificity

Epitope competition
In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.
Other clonesOverlap in epitope recognition with REA656
cA2++
mab11++
357-167-24-
Cells were incubated with an excess of purified unconjugated TNF-α (REA656) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison
Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.
View details
Flow cytometric comparison of different clones for TNF-α. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 20 ng/mL Phorbol 12-myristate 13-acetate (PMA) and 1 µg/ml Ionomycin for 1 hour, followed by an incubation with 1µg/mL brefeldin A for 4 hours. Cells were stained with a suitable counterstaining and then fixed and permeabilized followed by a staining with TNF-α antibodies. As a control, TNF-α antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD3+/CD4+ cells were pregated for the analysis. Cell debris and cell doublets were excluded from the analysis based on scatter signals. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for TNF-α. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 20 ng/mL Phorbol 12-myristate 13-acetate (PMA) and 1 µg/ml Ionomycin for 1 hour, followed by an incubation with 1µg/mL brefeldin A for 4 hours. Cells were stained with a suitable counterstaining and then fixed and permeabilized followed by a staining with TNF-α antibodies. As a control, TNF-α antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD3+/CD4+ cells were pregated for the analysis. Cell debris and cell doublets were excluded from the analysis based on scatter signals. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for TNF-α. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 20 ng/mL Phorbol 12-myristate 13-acetate (PMA) and 1 µg/ml Ionomycin for 1 hour, followed by an incubation with 1µg/mL brefeldin A for 4 hours. Cells were stained with a suitable counterstaining and then fixed and permeabilized followed by a staining with TNF-α antibodies. As a control, TNF-α antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD3+/CD4+ cells were pregated for the analysis. Cell debris and cell doublets were excluded from the analysis based on scatter signals. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for TNF-α. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 20 ng/mL Phorbol 12-myristate 13-acetate (PMA) and 1 µg/ml Ionomycin for 1 hour, followed by an incubation with 1µg/mL brefeldin A for 4 hours. Cells were stained with a suitable counterstaining and then fixed and permeabilized followed by a staining with TNF-α antibodies. As a control, TNF-α antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD3+/CD4+ cells were pregated for the analysis. Cell debris and cell doublets were excluded from the analysis based on scatter signals. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Flow cytometric comparison of different clones for TNF-α. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 20 ng/mL Phorbol 12-myristate 13-acetate (PMA) and 1 µg/ml Ionomycin for 1 hour, followed by an incubation with 1µg/mL brefeldin A for 4 hours. Cells were stained with a suitable counterstaining and then fixed and permeabilized followed by a staining with TNF-α antibodies. As a control, TNF-α antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD3+/CD4+ cells were pregated for the analysis. Cell debris and cell doublets were excluded from the analysis based on scatter signals. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Specifications for TNF-α Antibody, anti-human, REAfinity™

Overview

Clone REA656 recognizes the human tumor necrosis factor alpha (TNF-α) antigen, which is a cytokine involved in inflammatory immune responses. TNF-α has different functions, it plays an important role in the regulation of immune cells and is a potent pyrogen causing fever and apoptotic cell death. Furthermore, the TNF intracellular domain form is able to induce interleukin 12 (IL-12) production in dendritic cells. TNF-α is secreted by activated CD4
+
T cells, monocytes, macrophages, natural killer (NK) cells, mast cells, eosinophils, and neutrophils.
Additional information: Clone REA656 displays negligible binding to Fc receptors.

Alternative names

Tumor necrosis factor, Cachectin, TNF-alpha, TNF-a, DIF-alpha, TNFa, TNFSF2

Detailed product information

Technical specifications

CloneREA656
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (I), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenTNF-α
Alternative names of antigenTumor necrosis factor, Cachectin, TNF-alpha, TNF-a, DIF-alpha, TNFa, TNFSF2
Molecular mass of antigen [kDa]26
Distribution of antigenT cells, monocytes, macrophages, NK cells, mast cells, eosinophils, neutrophils
Entrez Gene ID7124
RRIDAB_2751595, AB_2752014, AB_2751984, AB_2751641

References for TNF-α Antibody, anti-human, REAfinity™

Publications

  1. Friedmann, E. et al. (2006) SPPL2a and SPPL2b promote intramembrane proteolysis of TNFalpha in activated dendritic cells to trigger IL-12 production. Nat. Cell Biol. 8(8): 843-848
  2. Nedwin, G. E. et al. (1985) Human lymphotoxin and tumor necrosis factor genes: structure, homology and chromosomal localization. Nucleic Acids Res. 13(17): 6361-6373
  3. Nedospasov, S. A. et al. (1985) Tandem arrangement of genes coding for tumor necrosis factor (TNF-alpha) and lymphotoxin (TNF-beta) in the human genome. Cold Spring Harb. Symp. Quant. Biol. 51(1): 611-624

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