M302B

antibody from Invitrogen Antibodies

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

Western blot

Antibody data

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Validation data

Product number: M302B - Provider product page
Provider: Invitrogen Antibodies
Product name: TNF alpha Monoclonal Antibody (2TNF-H33), Biotin
Antibody type: Monoclonal
Antigen: Recombinant full-length protein
Description: M302B targets TNF Alpha in ELISA, and WB applications and shows reactivity with Human samples. The M303 antibody (clone 2TNF-H34A) has successfully been paired as the coating antibody in a sandwich ELISA with detection antibody M302B (biotinylated conjugate of clone 2TNG-H33). The M302B immunogen is recombinant human TNF alpha. M302B detects TNF Alpha which has a predicted molecular weight of approximately 26 kDa. The M302B TNFa antibody (biotinylated conjugate of clone 2TNF-H33) has successfully been paired as the detection antibody in a sandwich ELISA with capture antibody M303 (clone 2TNF-H34A) Typical dilutions for sandwich ELISA include 1 µg/mL for coating and 0.125 - 0.25 µg/mL for detection. Biotinylated antibody M302B (clone 2TNF-H33) and antibody M303 (clone 2TNF-H34A) have successfully been used in combination with recombinant TNFa protein STNFA in ELISA applications.
Reactivity: Human
Host: Mouse
Conjugate: Biotin
Isotype: IgG
Antibody clone number: 2TNF-H33
Vial size: 250 μg
Concentration: 0.5 mg/mL
Storage: -20°C

TNF protein structure - M302B shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Sandwich ELISA analysis of Human TNF alpha Biotinylated MAb (Product # M302B) was performed using components of Thermo Fisher Scientific Human TNF alpha Colorimetric ELISA Kit (Product # EH3TNFA) by loading 50 µL per well of Recombinant Human TNF alpha in octuplicate at 1000, 500, 250, 125, 62.5, 31.2, 15.6, and 0 pg/mL across a 5 µg/mL rabbit anti-Human TNF pre-coated plate and incubated for 1 hour at room temperature. The plate was washed then incubated with 100 µl per well of Human TNF alpha Biotinylated MAb (Product # M302B) in duplicate at 2, 1, 0.5, and 0.25 µg/mL for 1 hour at room temperature. The plate was washed and incubated with 100 µl per well of Ultra Streptavidin-HRP (Product # N504) in all test wells at 1:1, 500 for 30 minutes at room temperature. The plate was washed then incubated with 100 µL per well of TMB substrate for 30 minutes at room temperature in the dark. The plate was then stopped with 0.16M sulfuric acid and absorbance was read on a spectrophotometer at 450-550 nm.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Sandwich ELISA analysis of Human TNF alpha Biotinylated MAb (Product # M302B) was performed using components of Thermo Fisher Scientific Human TNF alpha Colorimetric ELISA Kit (Product # EH3TNFA) by loading 50 µL per well of Recombinant Human TNF alpha in octuplicate at 1000, 500, 250, 125, 62.5, 31.2, 15.6, and 0 pg/mL across a 5 µg/mL rabbit anti-Human TNF pre-coated plate and incubated for 1 hour at room temperature. The plate was washed then incubated with 100 µl per well of Human TNF alpha Biotinylated MAb (Product # M302B) in duplicate at 2, 1, 0.5, and 0.25 µg/mL for 1 hour at room temperature. The plate was washed and incubated with 100 µl per well of Ultra Streptavidin-HRP (Product # N504) in all test wells at 1:1, 500 for 30 minutes at room temperature. The plate was washed then incubated with 100 µL per well of TMB substrate for 30 minutes at room temperature in the dark. The plate was then stopped with 0.16M sulfuric acid and absorbance was read on a spectrophotometer at 450-550 nm.
Conjugate: Biotin