Vector sg peroxidase substrate

Manufactured by Vector Laboratories

Vector® SG Peroxidase substrate is a chromogenic substrate for the detection of peroxidase enzyme activity. It produces a blue-green colored reaction product upon oxidation by peroxidase.

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Lab products found in correlation

Streptavidin peroxidase, by Agilent Technologies (1 mentions) Abc kit, by Agilent Technologies (1 mentions)

Close spelling variants of this product

Vector SG (15 citations) Vector SG substrate (4 citations) SG substrate (4 citations) Vector SG Peroxidase (HRP) Substrate Kit (4 citations) Vector SG Peroxidase substrate kit (4 citations) SG peroxidase substrate (3 citations) Biotinylated anti-rabbit IgG and Vector SG substrate kit peroxidase (2 citations)

2 protocols using vector sg peroxidase substrate

Quantifying Myocardial Tropoelastin and Macrophages

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Following the MRI scans, anesthetized mice were culled by neck dislocation and hearts were collected for ex vivo analysis (n=4 MI mice per time point, and n=3 SHAM-operated animals per time point). Hearts were harvested, the atriums were removed and the ventricles were washed in saline solution followed by immersion in 10% formaldehyde solution for 24h at room temperature. Hearts were then dehydrated, paraffin-embedded and transversely sectioned (5μm thick).
Immunohistochemistry (IHC) was used to quantify the amount of tropoelastin and macrophages in the myocardium. Tropoelastin was detected with anti-mouse rabbit polyclonal antibody (21600, Abcam; dilution 1:100) using an avidin-biotin-peroxidase method (Vector® SG Peroxidase substrate; Vector Laboratories, Burlingame, CA). A monoclonal rat anti-mouse antibody (550292, BD Pharmingen) (CD107b; MAC-3; dilution 1:100) was used for macrophage detection. The antibody was revealed with streptavidin-peroxidase (Dako, Ely, UK) (ABC kit, 1:100). Digital images were analyzed using ImageJ (National Institute of Health, Bethesda, MD). Tropoelastin and MAC-3 were quantified and expressed as percentage of the infarcted myocardium using ImageJ, and were manually segmented and normalized with the total area of infarction for each histology slice calculated from Masson’s trichrome staining.

Ramos I.T., Henningsson M., Nezafat M., Lavin B., Lorrio S., Gebhardt P., Protti A., Eykyn T.R., Andia M.E., Flögel U., Phinikaridou A., Shah A.M, & Botnar R.M. (2018). Simultaneous Assessment of Cardiac Inflammation and Extracellular Matrix Remodeling after Myocardial Infarction. Circulation. Cardiovascular imaging, 11(11), e007453.

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Immunohistochemical Staining Protocol

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Endogenous peroxidase was inhibited by incubation of the slides in 3% oxygenated water and methanol for 10 minutes. Slides were then washed in deionized water for 10 minutes. Antigen retrieval was performed by boiling the samples for 2 minutes at 110 °C in citrate buffer in a pressurized antigen retrieval unit and then allowed to cool. Samples were washed with PBS for 3 minutes, and blocking was performed for 1 hour at room temperature with 10% donkey serum in PBS. Incubation with the primary antibody was carried out overnight at 4 ˚C diluted 1:100 in PBS 2% normal donkey serum. Samples were allowed to temper at room temperature for 1 hour, then slides were washed twice in PBS for 20 minutes. Slides were incubated with anti-rabbit HRP polymer for 45 minutes at room temperature then washed 3 times in PBS for 5 minutes.
Slides were revealed with Vector SG Peroxidase substrate for 10 minutes at room temperaturePBS (5 ml), Chromogen (75 μl), Hydrogen peroxide (120 μl).
Samples were washed twice in PBS for 5 minutes, before counterstaining with nuclear fast red for 5 minutes. Slides were washed with deionized water then finally dehydrated through a gradient of ethanol, cleared in xylene, and mounted.

Evans R.J., Lavin B., Phinikaridou A., Chooi K.Y., Mohri Z., Wong E., Boyle J.J., Krams R., Botnar R, & Long N.J. (2020). Targeted Molecular Iron Oxide Contrast Agents for Imaging Atherosclerotic Plaque. Nanotheranostics, 4(4), 184-194.

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