Clone:
cA2
Type of antibody:
Primary antibodies, Functional-grade antibodies
Isotype:
human IgG1
Applications:
ICFC, MICS, IF, IHC, FA
Alternative names:
TNF, DIF, TNFSF2

Extended validation for TNF-α Antibody, anti-human

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 cA2
REA656++
mab11++
357-167-24-
Cells were incubated with an excess of purified unconjugated TNF-α (cA2) 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

Overview

Tumor necrosis factor alpha (TNF-α) is produced by cells that are involved in inflammatory immune responses. TNF-α is secreted by activated CD4
+
T cells, monocytes, macrophages, NK cells, and neutrophils.
Anti-TNF-α antibodies have been designed for intracellular staining of TNF-α–producing cells. Cells can be stimulated for TNF-α production, for example, by polyclonal stimulation with mitogens. For induction of TNF-α production by antigen-specific T cells, cells are restimulated with the respective antigen. TNF-α can be accumulated in the cells by addition of secretion inhibitors like brefeldin A. After fixation and permeabilization of the cell sample, TNF-α–producing cells can be stained intracellularly with Anti-TNF-α antibodies. Staining for surface markers allows simultaneous flow cytometric analysis of subsets and activation status of the TNF-α–producing cells.

Alternative names

TNF, DIF, TNFSF2

Detailed product information

Technical specifications

ClonecA2
Clonalitymonoclonal
Isotypehuman IgG1
Hosthuman
Type of antibodyPrimary antibodies, Functional-grade antibodies
Specieshuman
Cross-reactivity
rhesus monkey (
Macaca mulatta
)
,
cynomolgus monkey (
Macaca fascicularis
)
AntigenTNF-α
Alternative names of antigenTNF, DIF, TNFSF2
Molecular mass of antigen [kDa]26
Entrez Gene ID7124
RRIDAB_2752116, AB_2752165, AB_2752115, AB_2751396, AB_2751384, AB_2784484, AB_2784483, AB_2752167, AB_2752117, AB_10839558, AB_2752166

Resources for TNF-α Antibody, anti-human

Certificates

Please follow this
link
to search for Certificates of Analysis (CoA) by lot number.

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