Immunolon 2HB 96 well flat bottom microtiter plates (Thermo Scientific, Waltham, MA, USA) were coated with unlabeled Ig (1010-01) (Southern Biotech, Birmingham, AL, USA) and blocked the following day with 5% gelatin in 1x PBS. Serum samples were diluted to 1:100,000 for IgM and IgG, 1:50,000 for IgA, IgG1, and IgG3, and 1:100,000 for IgG2b and IgG2c and plated in duplicate. HRP-conjugated secondary antibodies specific to IgG (cat#: 1032-05), IgM (cat#: 1020-05), IgA (cat#: 1040-05) IgG1 (cat#: 1070-05), IgG2b (cat#: 1090-05), IgG2c (cat#: 1079-05), and IgG3 (cat#: 1100-05) (Southern Biotech, Birmingham, AL, USA) were added. Plates were developed with 3,3′,5,5′-Tetramethylbenzidine (TMB) substrate kit (Thermo Scientific, Waltham, MA, USA) and absorbance OD450nm was read using a Victor 3 plate reader (Perkin Elmer, Waltham, MA, USA).
3 3 5 5 tetramethylbenzidine tmb substrate kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The 3,3′,5,5′-Tetramethylbenzidine (TMB) substrate kit is a laboratory reagent used as a colorimetric substrate for various enzyme-linked assays, such as ELISA. The kit provides the necessary components to prepare a TMB-based solution that can be used to detect and quantify the presence of target analytes in a sample.
Automatically generated - may contain errors
Lab products found in correlation
96 well microplate, by SPL Life Sciences (2 mentions)
Isopropyl β d 1 thiogalactopyranoside (iptg), by Merck Group (2 mentions)
Igg2c, by Southern Biotech (1 mentions)
Igg2b, by Southern Biotech (1 mentions)
Victor3 plate reader, by PerkinElmer (1 mentions)
Glycine, by Merck Group (1 mentions)
Hydrochloric acid (hcl), by Merck Group (1 mentions)
Tween 20, by Merck Group (1 mentions)
C reactive protein (crp), by Merck Group (1 mentions)
Xhoi, by New England Biolabs (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Lysozyme, by Merck Group (1 mentions)
Sucrose, by Merck Group (1 mentions)
3 protocols using 3 3 5 5 tetramethylbenzidine tmb substrate kit
1
Multiplex Antibody Isotyping ELISA
2
Immunoassay Reagents and Buffers
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Horseradish peroxidase (HRP), C-reactive protein (CRP), human IgG (hIgG), Tween-20, glycine, Isopropyl β-d -1-thiogalactopyranoside (IPTG), ethylenediaminetetraacetic acid (EDTA), SDS and hydrochloric acid were obtained from Merck (Darmstadt, Germany). Rabbit polyclonal anti-HRP antibodies, rabbit polyclonal anti-CRP antibodies, fluorescein isothiocyanate conjugated goat polyclonal anti-CRP (anti-CRP(FITC)) antibodies, and fluorescein isothiocyanate labeled goat polyclonal anti-human (anti-human(FITC)) antibodies were purchased from Abcam (Cambridge, UK). Phosphate buffered saline (PBS) was obtained from LPS solution (Daejeon, Korea) and used as antibody dissolving buffer. In addition, 96-well microplates were purchased from SPL Life Science (Pocheon, Korea). The 3,3′,5,5′-Tetramethylbenzidine (TMB) substrate kit was obtained from Thermo Fisher Scientific (Waltham, MA, USA).
3
Immunoassay Reagents and Materials
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Goat polyclonal antibodies against C-reactive protein (CRP), goat polyclonal antibodies against horseradish peroxidase (HRP), goat polyclonal antibodies against CRP labeled with HRP, and fluorescein-conjugated rabbit polyclonal antibodies against human immunoglobulin (Ig) G were obtained from Abcam (Cambridge, UK). Tris-HCl buffer ( pH 8.0), Tris-glycine buffer, phosphate-buffered saline (PBS), Luria-Bertani (LB) broth, ampicillin, protein G, and sodium dodecyl sulfate (SDS)-loading buffer were purchased from LPS solution (Daejeon, Korea). 96-Well microplates were obtained from SPL Life Sciences (Pocheon, Korea). The 3,3′,5,5′-tetramethylbenzidine (TMB) substrate kit was purchased from Thermo Fisher Scientific (Waltham, MA, USA). Triton X-100, isopropyl β-D-1-thiogalactopyranoside (IPTG), sucrose, lysozyme, HRP, CRP, and other analytical-grade chemicals were obtained from Merck (Darmstadt, Germany). The restriction enzymes, XhoI and KpnI, and ligase were purchased from New England Biolabs (Ipswich, MA, US).
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