Pierce streptavidin poly hrp

Manufactured by Thermo Fisher Scientific

The Pierce™ streptavidin poly-HRP is a reagent used in laboratory applications. It consists of streptavidin, a protein that binds to biotin, conjugated with multiple horseradish peroxidase (HRP) enzymes. This product can be used in various immunoassay and detection techniques that involve biotin-streptavidin interactions.

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Lab products found in correlation

Xyz 3060, by BioDot (1 mentions) Goat anti mouse igg, by Jackson ImmunoResearch (1 mentions) Total tau antibody, by Agilent Technologies (1 mentions) Startingblock blocking buffer, by Thermo Fisher Scientific (1 mentions) Nunc maxisorp plate, by Thermo Fisher Scientific (1 mentions) Sodium chloride (nacl), by Merck Group (1 mentions) Ethylene glycol tetraacetic acid (egta), by Merck Group (1 mentions) Mgcl2, by Merck Group (1 mentions) 1 step ultra tmb elisa, by Thermo Fisher Scientific (1 mentions) Nunc polysorp 96 well plates, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Streptavidin-HRP (301 citations) Streptavidin Poly-HRP (14 citations)

4 protocols using pierce streptavidin poly hrp

Sandwich Immunoassay for Influenza NP Detection

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A schematic of the sandwich immunoassay implemented to detect NP in nitrocellulose (NC) and the process control are shown in Figure 1. The target nucleoprotein capture antibodies, which were type-specific for either influenza A or B (0.4 μL, 1 mg/mL, mouse monoclonal anti-influenza A/B NP IgG, Hytest Ltd), and a process control capture antibody (0.3 μL, 0.4 mg/mL, goat anti-mouse IgG, Jackson ImmunoResearch) were immobilized on 5-mm-wide NC membrane (Millipore, HF120) strips, and spaced 4 mm apart via printing with a reagent dispenser (BioDot, XYZ3060), followed by drying at 37°C for 2 hr in a box with desiccant. Then the strips were stored in a sealed pouch with desiccant overnight in a humidity-controlled room (20% RH); they were cut to final size with a Matrix™ 2360 programmable shear (Kinematic Automation, CA). Diaminobenzidine (DAB), in the presence of hydrogen peroxide (H₂O₂), is enzymatically converted from colorless substrate to a brown precipitate by horseradish peroxidase (HRP). The two detection antibodies, which were biotinylated and type-specific for either influenza A or B (biotinylated mouse monoclonal anti-influenza AB NP, Hytest Ltd) were conjugated with Pierce™ streptavidin poly-HRP (Thermo Fisher Scientific, Waltham, MA), which contains 2-8 HRP molecules per streptavidin tetramer.

Huang S., Abe K., Bennett S., Liang T., Ladd P.D., Yokobe L., Anderson C.E., Shah K., Bishop J., Purfield M., Kauffman P.C., Paul S., Welch A.E., Strelitz B., Follmer K., Pullar K., Sanchez-Erebia L., Gerth-Guyette E., Domingo G., Klein E., Englund J.A., Fu E, & Yager P. (2017). Disposable Autonomous Device for Rapid Swab-to-Result Diagnosis of Influenza. Analytical chemistry, 89(11), 5776-5783.

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Tau Protein Detection Assay

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The coating buffer containing 1 μg/mL of total tau antibody (DAKO) in carbonate-bicarbonate buffer (pH 9.6) was pipetted into a 96-well plate (100 μL per well) and left overnight at 4 °C. The wells were washed three times with TBST (Tris-buffered saline, with Tween 20, pH 8.0, Sigma) and incubated with StartingBlock Blocking Buffers (Thermo Scientific) for 1 h at room temperature. After removal of the blocking buffer and washing three times with TBST, different concentrations of Tau441 protein were added into each well (100 μL). The plate was stored at 4 °C overnight.
On the next day, the plate was washed three times with TBST and then treated with 500 nM detecting aptamer or randomly scrambled aptamer sequence (RS) in 100 μL of PVP buffer (20 mg/mL polyvinylpyrrolidone and 100 μg/mL salmon sperm in TBST). The reaction was left on a shaker for 1 h. Afterward, the wells were washed with TBST three times and 100 μL of 1:10K diluted Pierce Streptavidin Poly-HRP (Thermo Scientific) solution was added to each well and incubated for 1 h at room temperature. Finally, 100 μL of TMB solution was added to each well, and the 620 nm absorbance was recorded by a plate reader.

Teng I.T., Li X., Yadikar H.A., Yang Z., Li L., Lyu Y., Pan X., Wang K.K, & Tan W. (2018). Identification and Characterization of DNA Aptamers Specific for Phosphorylation Epitopes of Tau Protein. Journal of the American Chemical Society, 140(43), 14314-14323.

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Quantifying Complement Activation by ELISA

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Complement activation via the alternative pathway was measured by an enzyme-linked immunosorbent assay based on Seelen et al. [43 (link)]. Briefly, a Nunc MaxiSorp plate (Thermo Scientific) was coated overnight with 1 µg/ml of LPS isolated from the WT and S. Enteritidis PCM 2817 mutants in PBS containing 10 mM MgCl₂ (Sigma–Aldrich). The plate was washed three times with PBS containing 0.05% Tween-20 (PBS-T). NHS dilutions were selected on the basis of preliminary tests and prepared in Tris-buffered saline (TBS; 10 mM Tris base and 150 mM NaCl (Sigma–Aldrich), pH 7.4) containing 5 mM MgCl₂, 10 mM EGTA (MgEGTA, Sigma–Aldrich), 1% w/v bovine serum albumin (BSA), 0.05% Tween-20 and incubated for 1 h at 37 °C. The plate was washed three times with PBS-T. Next, biotinylated anti-human C5b-9 neoepitope antibody (Hycult, clone aE11) in TBS containing 1% w/v BSA (B-TBS) was added to the wells and the plate was incubated for 1 h at 37 ℃. After washing, streptavidin conjugated with horseradish peroxidase (Pierce^™ Streptavidin Poly-HRP, Thermo Scientific) in B-TBS was added and incubated for 1 h at 37 °C. After washing, the reaction was revealed with 3,3′,5,5′-tetramethylbenzidine (TMB) substrate, stopped with 0.5 M sulphuric acid, and absorbance read at 450 nm. Absorbance readings were normalized to a wild-type value of 100%. The experiment was performed in three biological replicates.

Krzyżewska-Dudek E., Dulipati V., Kapczyńska K., Noszka M., Chen C., Kotimaa J., Książczyk M., Dudek B., Bugla-Płoskońska G., Pawlik K., Meri S, & Rybka J. (2024). Lipopolysaccharide with long O-antigen is crucial for Salmonella Enteritidis to evade complement activity and to facilitate bacterial survival in vivo in the Galleria mellonella infection model. Medical Microbiology and Immunology, 213(1), 8.

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Magnetic Bead-based IgG Capture ELISA

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Tosyl-activated superparamagnetic microbeads M-450 (MBs, Dynabeads, Cat. no. 14013, 4.5 μm diameter, 4 × 10⁸ beads/mL, 30 mg/mL) were from Invitrogen. TDM from Mycobacterium bovis (Cat. no. T3034), and fatty acid-free bovine serum albumin (BSA, Cat. no. A7030) were from Sigma. Goat F(ab’)₂ anti-human IgG (H + L) labelled with biotin was from Southern Biotech. Nunc PolySorp 96-well plates, 1-step ultra TMB ELISA, and Pierce streptavidin poly-HRP were obtained from Thermo Scientific. Mycobacterial recombinant 38 kDa and Ag85A proteins were obtained from Mybiosource (CA, USA). Neodymium disc magnets (diameter 5 mm, thickness 2 mm) were from AliExpress Global Retail. Unless otherwise specified, all experiments were performed using PBS buffer without Ca⁺² and Mg⁺² ions.

Mani V., Paleja B., Larbi K., Kumar P., Tay J.A., Siew J.Y., Inci F., Wang S., Chee C., Wang Y.T., Demirci U., De Libero G, & Singhal A. (2016). Microchip-based ultrafast serodiagnostic assay for tuberculosis. Scientific Reports, 6, 35845.

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