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Product Name
Anti-TNFRSF18 antibody
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Description
Mouse monoclonal antibody to TNFRSF18
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Tested applications
WB, IHC-P, FC
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Species reactivity
Human
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Alternative names
AITR antibody; GITR antibody; CD357 antibody; GITR-D antibody; ENERGEN antibody
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Isotype
Mouse IgG1
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Preparation
This antigen of this antibody was purified recombinant fragment of human tnfrsf18 (aa: extra 26-162) expressed in e. coli.
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Clonality
Monoclonal
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Formulation
Liquid, 1*PBS with 0.05% sodium azide.
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Storage instructions
Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
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Applications
WB: 1:500-1:2,000
IHC-P: 1:50-1:200
FC: 1:100-1:200
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Validations
Fig1: Western blot analysis of TNFRSF18 against human TNFRSF18 (AA: extra 26-162) recombinant protein. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody ( 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody at 1:5,000 dilution was used for 1 hour at room temperature.
Fig2: Western blot analysis of 175074# against HEK293 (1) and TNFRSF18 (AA: extra 26-162)-hIgGFc transfected HEK293 (2) cell lysate.Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody ( 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody at 1:5,000 dilution was used for 1 hour at room temperature.
Fig3: Immunohistochemical analysis of paraffin-embedded human cervical cancer tissue using anti-TNFRSF18 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody ( 1/100) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Fig4: Immunohistochemical analysis of paraffin-embedded human colon cancer tissue using anti-TNFRSF18 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody ( 1/100) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Fig5: Flow cytometric analysis of TNFRSF18 was done on Jurkat cells. The cells were fixed, permeabilized and stained with the primary antibody ( 1/100) (green). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated goat anti-Mouse IgG Secondary antibody at 1/500 dilution for 30 minutes. Unlabelled sample was used as a control (cells without incubation with primary antibody; red).
- Background
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References
- Immunol Lett. 2014 Dec;162(2 Pt B):210-6.
- Int J Rheum Dis. 2016 Feb;19(2):199-204.
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