Clone:
REA636
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
ICFC
Alternative names:
TNFa, TNFalpha, TNF-alpha, Cachectin, Tnfsf1a, TNFSF2

Extended validation for TNF-α Antibody, anti-mouse, 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 REA636
MP6-XT22++
TN3-19.12-
Cells were incubated with an excess of purified unconjugated TNF-α (REA636) 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. Splenocytes from Balbc/c mice were stimulated with 50 ng/mL CD3 + 1 µg/mL CD28 for 2h, followed by an incubation with 1µg/mL brefeldin A for 4 hours. Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Set followed by intracellular staining with TNF-αantibodies. 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. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. 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. Splenocytes from Balbc/c mice were stimulated with 50 ng/mL CD3 + 1 µg/mL CD28 for 2h, followed by an incubation with 1µg/mL brefeldin A for 4 hours. Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Set followed by intracellular staining with TNF-αantibodies. 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. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. 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. Splenocytes from Balbc/c mice were stimulated with 50 ng/mL CD3 + 1 µg/mL CD28 for 2h, followed by an incubation with 1µg/mL brefeldin A for 4 hours. Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Set followed by intracellular staining with TNF-αantibodies. 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. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. 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. Splenocytes from Balbc/c mice were stimulated with 50 ng/mL CD3 + 1 µg/mL CD28 for 2h, followed by an incubation with 1µg/mL brefeldin A for 4 hours. Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Set followed by intracellular staining with TNF-αantibodies. 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. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. 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. Splenocytes from Balbc/c mice were stimulated with 50 ng/mL CD3 + 1 µg/mL CD28 for 2h, followed by an incubation with 1µg/mL brefeldin A for 4 hours. Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Set followed by intracellular staining with TNF-αantibodies. 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. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

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

Overview

Clone REA636 recognizes the mouse tumor necrosis factor α (TNF-α) antigen, a 17 kDa cytokine that binds to TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. TNF-α is produced by cells that are involved in inflammatory immune responses. It is secreted by activated CD4
+
T cells, monocytes, macrophages, NK cells, and neutrophils. TNF-α binding to surface receptors elicits a wide array of biologic 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. Furthermore it causes fever by direct action or by stimulation of interleukin-1 secretion.
Additional information: Clone REA636 displays negligible binding to Fc receptors.

Alternative names

TNFa, TNFalpha, TNF-alpha, Cachectin, Tnfsf1a, TNFSF2

Detailed product information

Technical specifications

CloneREA636
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody, human IgG1
Hosthuman cell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Speciesmouse
AntigenTNF-α
Alternative names of antigenTNFa, TNFalpha, TNF-alpha, Cachectin, Tnfsf1a, TNFSF2
Molecular mass of antigen [kDa]26
Distribution of antigenT cells
Entrez Gene ID21926
RRIDAB_2819466, AB_2889792, AB_2784485, AB_2784486

Resources for TNF-α Antibody, anti-mouse, REAfinity™

Certificates

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

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

Publications

  1. Chyuan, I. T. et al. (2015) Tumor necrosis factor-alpha blockage therapy impairs hepatitis B viral clearance and enhances T-cell exhaustion in a mouse model. Cell. Mol. Immunol. 12(3): 317-325
  2. Torrente, Y. et al. (2003) Tumor necrosis factor-alpha (TNF-alpha) stimulates chemotactic response in mouse myogenic cells. Cell Transplant 12(1): 91-100
  3. Kaiser, G. C. et al. (1997) Tumor necrosis factor alpha regulates proliferation in a mouse intestinal cell line. Gastroenterology 112(4): 1231-1240

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