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Western blot
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- Antibody Data
- Antigen structure
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- Validations
- Other assay [1]
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- Product number
- M302 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- TNF alpha Monoclonal Antibody (2TNF-H33)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- Description
- The M303 anti-TNF Alpha antibody (clone 2TNF-H34A) has successfully been paired as the coating antibody in a sandwich ELISA with detection antibody M302B (biotinylated conjugate of clone 2TNF-H33). This product has been tested for endotoxins by limulus amoebocyte lysate (LAL) assay and contains an endotoxin concentration of less than or equal to 10 endotoxin units per milligram (EU/mg).
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 2TNF-H33
- Vial size
- 500 µg
- Concentration
- 1 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
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Supportive validation
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- Invitrogen Antibodies (provider)
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- Experimental details
- Direct ELISA analysis of TNF alpha was performed by coating wells of a 96-well plate with 100 µL per well of human recombinant protein TNF alpha (Product # RTNFAI) diluted in carbonate/bicarbonate buffer (Product # 28382) at a concentration of 1 µg/mL overnight at 4C. Wells of the plate were washed, blocked with starting blocking buffer (Product # 37538), and incubated with 100 µL per well of a mouse anti-TNF alpha monoclonal antibody (Product # M302) at a dilution of 1:50, 1:100, 1:200, 1:400, 1:800, 1:1600 and 1:3200 for 90 minutes at 37C. The plate was washed, then incubated with 100 µL per well of a HRP-conjugated goat anti-mouse IgG secondary antibody (Product # 62-6520) at a dilution of 1:2000 for 90 minutes at 37C. Detection was performed using 1-Step Ultra TMB substrate (Product # 34028) for 5-10 minutes at room temperature in the dark. The reaction was stopped with Stop solution (Product # N600), and absorbances were read on a spectrophotometer at 450-550 nm.
