Tmb substrate set

Manufactured by BD

The TMB Substrate Set is a laboratory product used for the detection and quantification of horseradish peroxidase (HRP) in enzyme-linked immunosorbent assay (ELISA) applications. The set contains the necessary components to prepare a TMB (3,3',5,5'-Tetramethylbenzidine) substrate solution, which undergoes a color change in the presence of HRP, allowing for the measurement of HRP activity.

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Lab products found in correlation

Streptavidin hrp, by BD (3 mentions) Nunc maxisorp 96 well plate, by Thermo Fisher Scientific (2 mentions) Human il 6 elisa kit, by BD (1 mentions) Ifn γ, by Thermo Fisher Scientific (1 mentions) Recombinant il 4, by R&D Systems (1 mentions) Il 13, by Thermo Fisher Scientific (1 mentions) Ifn γ, by R&D Systems (1 mentions) Il 13, by R&D Systems (1 mentions) Biotinylated anti areg antibody, by R&D Systems (1 mentions) Ova grade 5, by Merck Group (1 mentions) Anti mouse ige, by BD (1 mentions) Purified mouse igg1, by BD (1 mentions) Purified mouse ige, by BD (1 mentions)

Close spelling variants of this product

TMB substrate (104 citations) TMB Substrate Reagent Set (77 citations) BD OptEIA TMB substrate reagent set (13 citations) BD OptEIA TMB substrate set (3 citations) 3,3′,5,5′-tetramethylbenzidine (TMB) substrate reagent set (2 citations)

4 protocols using tmb substrate set

Quantifying IL-6 in Senescent Cells

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Human IL-6 ELISA kit (Cat. 55522) and TMB substrate set (Cat. 555214) were purchased from BD Biosciences and used according to manufacturer’s instructions. Conditioned medium samples from senescent HF043 cells in a 384 well plate at ~ 1000 cells per well were diluted fivefold in DMEM-FBS prior to assay.

Rolt A., Nair A, & Cox L.S. (2019). Optimisation of a screening platform for determining IL-6 inflammatory signalling in the senescence-associated secretory phenotype (SASP). Biogerontology, 20(3), 359-371.

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Cytokine Quantification in Biological Samples

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Supernatants collected from re-stimulated lymph node cells or BAL samples were added to Nunc Maxisorp 96-well plates (Thermo Fisher Scientific), which had been coated with purified anti-mouse IL-4 antibody (Thermo Fisher Scientific), IL-5 (BD Biosciences), IL-13 (Thermo Fisher Scientific) and IFN-γ (Thermo Fisher Scientific) all at 2 μg/ml O/N at 4 °C. Plates were subsequently blocked with 1% BSA. Cytokines were detected with biotinylated anti-mouse IL-4 (Thermo Fisher Scientific), IL-5 (Thermo Fisher Scientific), IL-13 (Thermo Fisher Scientific) and IFN-γ (Thermo Fisher Scientific) all at 0.5 μg/ml. Recombinant IL-4, IL-5, IL-13 and IFN-γ (R&D systems) were used as a standard (starting concentration 10 ng/ml). Antibody binding was detected with streptavidin-HRP (BD Biosciences) and developed with TMB substrate set (BD Biosciences).

Agaronyan K., Sharma L., Vaidyanathan B., Glenn K., Yu S., Annicelli C., Wiggen T.D., Penningroth M.R., Hunter R.C., Dela C.S, & Medzhitov R. (2022). Tissue remodeling by an opportunistic pathogen triggers allergic inflammation. Immunity, 55(5), 895-911.e10.

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Quantification of Areg in Human and Mouse Samples

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Human lung H292 cells were plated and stimulated as indicated in respective figures in 24-well plates at 1.5 × 105 cells per well. 300–500 μl of cell supernatant was collected and frozen at −20 °C until further use. Mouse BAL samples were used for mouse Areg ELISA. For Sandwich ELISA Nunc Maxisorp 96-well plates (Thermo Fisher Scientific) were coated with capture human Areg antibody (R&D systems) at 5 μg/ml or mouse Areg antibody (R&D systems) at 0.5 μg/ml and incubated O/N at 4 °C. Plates were washed with PBS-T and blocked with 1 % BSA for 1hr at RT. Human Areg was detected with biotinylated anti-Areg antibody (R&D systems). Mouse Areg was detected with biotinylated anti-Areg antibody (R&D systems). Antibody binding was detected with streptavidin-HRP (BD Biosciences) and developed with TMB substrate set (BD Biosciences). Recombinant human (GeneScript) and mouse Areg (BioLegend) were used as standards.

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Quantifying Murine Antibody Responses

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For serum OVA-specific IgG1 antibody detection, Nunc Maxisorp 96-well plates (Thermo Fisher Scientific) were coated with 10 μg/ml OVA (Grade V, Sigma-Aldrich) O/N at 4°C, subsequently blocked with 1 % BSA and serial dilutions of sera were added (1:1000, 1:2500, 1:5000, 1:10000). Purified mouse IgG1 (BD Biosciences) was used as standard (starting concentration 500 ng/ml). Detection was achieved with biotinylated anti-IgG1 (BD Biosciences) at 0.1 μg/ml. For total IgE detection, the plates were coated with anti-mouse IgE (BD Biosciences) at 2 μg/ml and antibody was detected with biotinylated anti-IgE (BD biosciences) at 0.1 μg/ml. Purified mouse IgE (BD Biosciences) was used as standard (starting concentration 500 ng/ml). For OVA-specific IgE detection, the plates were coated with anti-mouse IgE (BD Biosciences) as above and the antibody was detected by biotin-OVA (Nanocs) at 10 μg/ml. Mouse anti-OVA IgE (Bio-Rad was used as a standard (starting concentration 250 ng/ml). Antibody binding was detected with streptavidin-HRP (BD Biosciences) and developed with TMB substrate set (BD Biosciences).

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