- MA1-116 - Invitrogen Antibodies TGFB1 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of human TGF beta-1 was performed by loading 10 µL of human serum from patients with multiple sclerosis (MS), leukemia, or asthma (left panel) and 2 µg and 1 µg of recombinant human TGF beta-1 (right panel) protein per well on a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with StartingBlock (TBS) Blocking Buffer (Product # 37542) for at least 1 hour. The membrane was probed with a TGF beta-1 monoclonal antibody (Product # MA1-116) at a concentration of 5 µg/mL overnight at 4C on a rocking platform, washed in TBS-0.1%Tween-20, and probed with a goat anti-mouse IgG-HRP secondary antibody (Product # 31430) at a dilution of 1:10,000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34075).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Sandwich ELISA analysis of an anti-human TGF beta-1 monoclonal antibody (Product # MA1-116) was performed by loading 100 µL per well of coating antibody (Product # MA1-169) at a concentration of 2 µg/mL overnight at room temperature. The plate was washed 3 times with ELISA Wash Buffer (Product # N503), and 100 µL of recombinant TGF beta-1 was added to wells in duplicate at 2.5, 1.25, 0.625, 0.312, 0.156, 0.078, 0.039 and 0 ng/mL concentrations and the samples were incubated for 2 hours at room temperature. The plate was washed, then incubated with 100 µL per well of a biotinylated TGF beta-1 monoclonal antibody (Product # MA1-116), biotinylated using EZ-Link Sulfo-NHS-LC-Biotinylation Kit (Product # 21435), at a concentration of 0.125 µg/mL for 1 hour at room temperature, followed by 100 µL per well of Streptavidin-HRP (Product # N504) at a dilution of 1:40,000 for 30 minutes at room temperature. Detection was performed by adding 100 µL of 1-Step Ultra TMB substrate (Product # 34028) per well and incubating for 20 minutes at room temperature in the dark. The plate was then stopped with 100 µL per well of 0.16M sulfuric acid. Absorbances were read on a spectrophotometer at 450-550 nm.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Direct ELISA analysis of an anti-TGF beta-1 monoclonal antibody (Product # MA1-116) was performed by coating wells of a plate with recombinant TGF beta-1 protein at a concentration of 2 µg/mL overnight at 4C. The plate was washed 3 times with ELISA Wash Buffer (Product # N503), and 100 µL of a TGF beta-1 monoclonal antibody (Product # MA1-116) was added to wells in duplicate at 2000, 1000, 500, 250, 125, 62.5, 31.25, 15.6, 7.8, 3.9, 1.9, and 0 ng/mL concentrations and the samples were incubated for 2 hours at room temperature. The plate was washed then incubated with 100 µL per well of an HRP-conjugated goat anti-mouse IgG secondary antibody (Product # 31430) at a dilution of 1:10,000 for 1 hour at room temperature, and washed again with ELISA Wash Buffer. The plate was developed by incubating 100 µL per well of 1-Step Ultra TMB substrate (Product # 34028) per well and incubating for 10 minutes at room temperature in the dark. The reaction was stopped with 100 µL per well of 0.16M sulfuric acid. Absorbances were read on a spectrophotometer at 450-550 nm.

MA1-116