With gentle stirring, brain sections from the bregma level of mice exposed to 90 min MCAo were rinsed for 10 min at room temperature in 0.1 M PBS to remove intravascular IgG, and were fixed in 4% PFA [30 (link)]. Following the blocking of endogenous peroxidase with methanol/0.3% H2O2 and immersion in 0.1 M PBS containing 5% bovine serum albumin (BSA) and normal swine serum (1:1000), sections were incubated for 1 h in biotinylated goat anti-mouse IgG (sc-2013; Santa Cruz Biotechnology), and stained with an avidin peroxidase kit (Vectastain Elite; Vector Labs) and diaminobenzidine (Sigma). For reasons of data comparability, all sections were processed in parallel. Sections were scanned and IgG extravasation in the ischemic striatum and cortex was densitometrically analyzed. For correction of background staining, optical densities in corresponding contralateral non-ischemic tissue were subtracted from those in the ischemic tissue.
Biotinylated goat anti mouse igg sc 2013
Manufactured by Santa Cruz Biotechnology
Biotinylated goat anti-mouse IgG (sc-2013) is a laboratory reagent used for the detection and quantification of mouse immunoglobulin G (IgG) in various experimental applications. It is a secondary antibody conjugated with biotin, which can be used in conjunction with streptavidin-based detection systems.
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2 protocols using biotinylated goat anti mouse igg sc 2013
1
Quantifying Ischemic IgG Extravasation
2
Immunohistochemical Analysis of IgG Extravasation
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Brain sections at the level of bregma of mice exposed to 90 min MCAo, were rinsed for 10 min at room temperature in 0.1 M PBS to remove intravascular IgG, and were fixed in 4% PFA [5] (link). Following the blocking of endogenous peroxidase with methanol/0.3% H2O2 and immersion in 0.1 M PBS containing 5% bovine serum albumin (BSA) and normal swine serum (1:1000), sections were incubated for 1 h in biotinylated goat anti-mouse IgG (sc-2013; Santa Cruz Biotechnology), and stained with an avidin peroxidase kit (Vectastain Elite; Vector Labs) and diaminobenzidine (Sigma). All sections were processed in parallel for standardization. Sections were scanned and IgG extravasation in the ischemic striatum and cortex was densitometrically analyzed. For correction of background staining, optical densities in corresponding contralateral non-ischemic tissue were subtracted from those in the ischemic tissue.
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