The cell lysates were diluted with PBS and 0.1 mL diluted cell lysate was added into appropriate wells. After the plates were incubated at 37°C for 2 h, the lysate was removed and washed four times with 200 μL PBS/ well. Then, 100 μL of the appropriate primary antibodies (1:3000) were added to each well and the plate was incubated at 37°C for 1 h. After washing four times with PBS, 100 μL HRP-linked secondary antibody was added to each well and the plate was incubated at 37°C for 30 min. After washing four times with PBS, 100 μL of TMB Substrate (SolarBio) was added to each well and the plate was incubated at 37°C for 10 min, and at 25°C for 30 min. Then 100 μL of ELISA STOP Solution (SolarBio) was added to each well and the plate was shaken gently for a few seconds. Finally, the absorbance at 450 nm was read using a BioRad microplate reader.
Tmb substrate
TMB substrate is a chromogenic substrate used in enzyme-linked immunosorbent assays (ELISA) and other immunoassays. It undergoes a color change reaction when catalyzed by the enzyme horseradish peroxidase (HRP), which is commonly used as a reporter enzyme in these assays.
Lab products found in correlation
15 protocols using tmb substrate
Quantifying Osteocalcin and Alkaline Phosphatase
The cell lysates were diluted with PBS and 0.1 mL diluted cell lysate was added into appropriate wells. After the plates were incubated at 37°C for 2 h, the lysate was removed and washed four times with 200 μL PBS/ well. Then, 100 μL of the appropriate primary antibodies (1:3000) were added to each well and the plate was incubated at 37°C for 1 h. After washing four times with PBS, 100 μL HRP-linked secondary antibody was added to each well and the plate was incubated at 37°C for 30 min. After washing four times with PBS, 100 μL of TMB Substrate (SolarBio) was added to each well and the plate was incubated at 37°C for 10 min, and at 25°C for 30 min. Then 100 μL of ELISA STOP Solution (SolarBio) was added to each well and the plate was shaken gently for a few seconds. Finally, the absorbance at 450 nm was read using a BioRad microplate reader.
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SARS-CoV-2 Spike Protein ELISA Assay
Binding Affinity of GLP-1 Fusion Proteins
ELISA for Serological Antibody Titers
Quantifying Angiogenic Factors by ELISA
SARS-CoV-2 Spike RBD Quantification
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