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TMB ELISA Peroxidase Substrate

Product Sizes
100 ml
TMBE-100-100ML
About this Product
SKU:
TMBE-100
Additional Names:
3,3',5,5'-Tetramethylbenzidine, TMB ELISA substrate, TMBE Substrate, chromogenic ELISA substrate
Application:
ELISA
CE/IVD:
RUO
Concentration:
1X
Extra Details:
TMB (3,3',5,5'-Tetramethylbenzidine) is a colorimetric substrate for peroxidase (HRP)-based enzyme immunoassays, notably ELISAs, and is known for producing a blue color that changes to yellow upon stopping the reaction with sulfuric acid. The color change is quantifiable at 450 nm. TMB's high sensitivity makes it ideal for detecting low concentrations of analytes.
Physical State:
Liquid - clear to very light blue colored liquid
Shipping Conditions:
Ambient
Specificity:
pH: 4.0 +/- 0.2 Stability @ 18° to 26°C: PASS Stability @ 4°C: PASS QC Raw Material: PASS Absorbance check of final product: PASS Performance Data per ELISA: PASS
Storage Conditions:
2-8[o]C unopened. Protect from light. Desiccated.
Supplier:
Rockland Inc
Type:
Proteins, Peptides, Small Molecules & Other Biomolecules: Substrates
Rockland Immunochemicals produces a wide variety of buffers and substrates for use in ELISAs. Antigen was diluted in ELISA Microwell Coating Stabilizer (p/n MB-063-0100) added to the microwell plate and incubated overnight at 4°C. The plate was then blocked with ELISA Microwell Blocking Buffer with Stabilizer (p/n MB-064-1000) for 2 hours. The primary antibody was diluted in PBS Fish Gel Concentrate (1:10)(p/n MB-066-0100), added to the plate, and allowed to incubate 1 hour at room temperature. HRP conjugated secondary antibody was diluted in HRP Conjugate Stabilizer (p/n MB-060-0100), added to the plate, and allowed to incubate for 30 minutes at room temperature. TMB ELISA Peroxidase Substrate (p/n TMBE-1000) was added to the plate and allowed to incubate for 30 minutes at room temperature. The reaction was then stopped with 1M HCl and read at 450nm.
ELISA results of Mouse Anti-SARS CoV-2 Spike Protein (S2) Antibody using TMBE-1000. Each well was coated in duplicate with 0.5ug of rec SARS CoV-2 Whole Spike protein [RED line], rec SARS CoV-2 Spike (S1) [GREEN line], rec SARS CoV-2 Spike (S1) RBD protein [BLUE line], and rec SARS CoV-2 Spike (S2) glycoprotein [PURPLE line]. The working dilution for SARS-CoV-2 whole spike is 1:190,000 and SARS CoV-2 Spike (S2) glycoprotein is 1:120,000. The starting dilution of antibody was 0.55ug/ml and the X-axis represents the Log10 of a 3-fold dilution. This titration is a 4-parameter curve fit where the IC50 is defined as the titer of the antibody. Assay performed using Rabbit Anti-Mouse IgG HRP conjugated (p/n 610-403-C46) at 1:8000 and TMB substrate (p/n TMBE-1000). SARS CoV-2 Spike (S2) EC50: 8ng/mL; SARS CoV-2 Whole Spike EC50: 5ng/mL.
TMB ELISA Peroxidase Substrate