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Bm blue pod substrate soluble

Manufactured by Roche

BM Blue POD Substrate soluble is a laboratory reagent used in colorimetric assays. It serves as a substrate for peroxidase enzymes, enabling the detection and quantification of these enzymes in various applications.

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3 protocols using bm blue pod substrate soluble

1

Serum Antibody Titer Determination by ELISA

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Example 5

Determination of Serum Titers (ELISA) from Immunized Rabbits

Human recombinant soluble TPBG extracellular domain was immobilized on a 96-well NUNC Maxisorp plate at 2 μg/ml, 100 μl/well, in PBS, followed by: blocking of the plate with 2% Crotein C in PBS, 200 μl/well; application of serial dilutions of antisera, in duplicates, in 0.5% Crotein C in PBS, 100 μl/well; detection with either (1) HRP-conjugated donkey anti-rabbit IgG antibody (Jackson Immunoresearch/Dianova), or (2) HRP-conjugated rabbit anti-human IgG antibody (Pierce/Thermo Scientific; 1/5000), or (3) biotinylated goat anti-human kappa antibody (Southern Biotech/Biozol; 1/5000) and streptavidin-HRP; each diluted in 0.5% Crotein C in PBS, 100 μl/well. For all steps, plates were incubated for 1 h at 37° C. Between all steps plates were washed 3 times with 0.05% Tween 20 in PBS. Signal was developed by addition of BM Blue POD Substrate soluble (Roche), 100 μl/well; and stopped by addition of 1 M HCl, 100 μl/well. Absorbance was read out at 450 nm, against 690 nm as reference. Titer was defined as dilution of antisera resulting in half-maximal signal.

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2

PDGF-B Binding Assay

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Example 23

Human recombinant PDGF-B was immobilized on a 96-well NUNC Maxisorb plate at 2.5 μg/ml (mouse) or 1.0 μg/ml (rabbit), 100 μl/well, in PBS, followed by: blocking of the plate with 2% CroteinC in PBS, 200 μl/well; application of serial dilutions of antisera, in duplicates, in 0.5% CroteinC in PBS, 100 μl/well; detection with HRP-conjugated goat anti-mouse IgG antibody (Jackson Immunoresearch) diluted 1:16,000 in 0.5% CroteinC in PBS for mouse sera or with biotinylated goat anti-human kappa antibody (Southern Biotech) diluted 1:5,000 and HRP-conjugated streptavidin diluted 1:8,000 in 0.5% CroteinC in PBS for rabbit sera, 100 μl/well. For all steps, plates were incubated for 1 hour at 37° C. Between all steps, plates were washed 3 times with 0.05% Tween 20 in PBS. Signal was developed by addition of BM Blue POD Substrate soluble (Roche Diagnostics GmbH, Mannheim, Germany), 100 μl/well; and stopped by addition of 1 M HCl, 100 μl/well. Absorbance was read out at 450 nm, against 690 nm as reference. Titer was defined as dilution of antisera resulting in half-maximal signal.

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3

ELISA for Anti-IL-1beta Antibody Titer

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Example 3

Human recombinant IL-1beta was immobilized on a 96-well NUNC Maxisorb plate at 2.5 μg/ml, 100 μl/well, in PBS, followed by: blocking of the plate with 2% CroteinC in PBS, 200 μl/well; application of serial dilutions of antisera, in duplicates, in 0.5% CroteinC in PBS, 100 μl/well; detection with HRP-conjugated goat anti-mouse IgG antibody (Jackson Immunoresearch) diluted 1:16,000 in 0.5% CroteinC in PBS, 100 μl/well. For all steps, plates were incubated for 1 hour at 37° C. Between all steps, plates were washed 3 times with 0.05 Tween 20 in PBS. Signal was developed by addition of BM Blue POD Substrate soluble (Roche Diagnostics GmbH, Mannheim, Germany), 100 μl/well; and stopped by addition of 1 M HCl, 100 μl/well. Absorbance was read out at 450 nm, against 690 nm as reference. Titer was defined as dilution of antisera resulting in half-maximal signal.

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