M302

antibody from Invitrogen Antibodies

Targeting: TNF DIF, TNF-alpha, TNFA, TNFSF2

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

M302

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).

Antibody clone number

2TNF-H33

Concentration

1 mg/mL

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

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.