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Alkaline phosphatase labeled goat anti rabbit igg antibody

Manufactured by Merck Group

Alkaline phosphatase-labeled goat anti-rabbit IgG antibody is a laboratory reagent used to detect and quantify rabbit immunoglobulin G (IgG) in various analytical techniques. The antibody is produced in goats and is conjugated with the enzyme alkaline phosphatase, which can be used to generate a colorimetric or chemiluminescent signal when exposed to the appropriate substrate.

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3 protocols using alkaline phosphatase labeled goat anti rabbit igg antibody

1

Quantifying Plasma C3c Complement Binding

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We used serial plasma dilutions (10%, 5%, 2.5%, 1.25%, and 0.75% in BBS) and pooled plasma of mice belonging to the same experimental group, incubating the samples for 15 min on mannan- (10 µg/mL) or acBSA-coated (25 µg/mL) plates prepared according to a procedure reported previously.18 (link) Plates were washed and incubated for 1 h 30 min at RT with a polyclonal anti-human-C3c antibody (Dako, A0062) diluted to 2.4 µg/mL in wash buffer. After washing, plates were incubated with an alkaline phosphatase-labeled goat anti-rabbit IgG antibody (Sigma A-3812) diluted to 1 µg/mL in wash buffer for 1 h 30 min at RT. After washing, the assay was developed by adding 100 µL of substrate solution (Sigma Fast p-Nitrophenyl Phosphate Tablets, Sigma). The absorbance at 405 nm was then measured using the Infinite M200 spectrofluorimeter managed by Magellan software (Tecan, CH).
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2

Quantifying Cell-Associated C3c Levels

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Fixed cells were washed three times with washing buffer (10 mM Tris-HCl, 140 mM NaCl, 5 mM CaCl2, 0.05% Tween 20, pH 7.4) and incubated with BSA (1% in washing buffer) for 2 h at RT. After three washings, cells were incubated with a rabbit polyclonal anti-human C3c (Dako, A0062) diluted 1:5000 in washing buffer, for 1 h 30 min at RT. After washing, cells were incubated with an alkaline-phosphatase-labeled goat anti-rabbit IgG antibody (Sigma A-3812) diluted 1:1000 in washing buffer for 1 h 30 min at RT. Cells were then washed and the assay was developed using substrate solution (Fast p-Nitrophenyl Phosphate tablets, Sigma). The absorption was read at OD 405 nm using the spectrofluorimeter (TECAN plate reader, Infinite M200, Switzerland).
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3

Cell Proliferation and Signaling Assays

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Sp was purchased from Sigma-Aldrich (St. Louis, MO, USA). 5-Bromo-2′-deoxyuridine (BrdU) Labeling reagent and the Detection Kit I were obtained from Roche Diagnostics GmbH (Mannheim, Germany). Anti-cyclin D1 and anti-p27 antibodies were from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). Anti-mitogen-activated protein kinase (MAPK), anti-PI3K p85 and anti-AKT antibodies were all purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). Alkaline phosphatase-labeled goat anti-rabbit IgG antibody and goat anti-mouse IgG antibody were both from Sigma-Aldrich.
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