Immunohistochemistry was performed to analyze the expression of BRG1 in the colon tissues according to standard protocols. The sections were heated at 60°C and deparaffinized. Then, the sections were placed in citric acid for antigen retrieval, and blocked with 5% BSA for 20 min, followed by stained with an anti-BRG1 antibody (1:200, Abcam, Cambridge, U. K) overnight at 4°C. The next day, the sections were observed with a microscope (Olympus Corporation, Japan).
Anti brg1 antibody
Manufactured by Abcam
Sourced in United Kingdom
Anti-BRG1 antibody is a laboratory reagent used to detect the presence and distribution of BRG1 protein in biological samples. BRG1 is a catalytic subunit of the SWI/SNF chromatin remodeling complex, which plays a crucial role in regulating gene expression. This antibody can be used in various applications, such as Western blotting, immunohistochemistry, and immunofluorescence, to study the expression and localization of BRG1 in cells and tissues.
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3 protocols using anti brg1 antibody
1
Immunohistochemical Analysis of BRG1 Expression
2
Quantitative Histological Profiling of Ly6G and BRG1
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Histological analyses were performed essentially as described before. Briefly, the paraffin embedded sections were blocked with 10% normal goat serum for 1 h at room temperature and then incubated with an anti‐Ly6G antibody (Abcam, cat# ab238132, 1:100) or anti‐BRG1 antibody (Abcam, cat# ab110641, 1:100). Staining was visualized by incubation with anti‐rabbit secondary antibody and developed with a streptavidin‐horseradish peroxidase kit (Pierce, cat# 21140) for 20 min. Pictures were taken using an Olympus IX‐70 microscope. Slides were observed under a light microscope at high power (X40) by two pathologists independently in a double‐blind fashion. The scoring system was based on the following criterion: the staining intensity was divided into quintiles; the slides with the strongest staining were given a score of 5 (top quintile) whereas the slides with the dimmest staining were given a score of 1 (bottom quintile).
3
ChIP Assay for BRG1 Binding to miR-148b Promoter
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For the chromatin immunoprecipitation (ChIP) assay, chromatin was prepared from A549 cells. A549 cells were treated with 10% formaldehyde for 10 minutes at room temperature, washed with phosphate buffered saline (PBS), and resuspended in lysis buffer (50 mM Tris-HCl pH 8.0, 40 mM ethylenediaminetetraacetic acid (EDTA), 0.5% SDS). Chromatin was prepared according to standard protocols and sheared to fragments in a Bioruptor (Diagenode, Philadelphia, PA, USA) by three sonication cycles of 5 minutes. The sheared chromatin was then immunoprecipitated by using anti-BRG1 antibody (Abcam, Cambridge, UK) or control IgG (Sigma, St. Louis, MO, USA). After extensive washing and elution, crosslinks were reverted by adjusting NaCl concentrations and heat treatment. Precipitated DNA fragments were purified by using PCR purification kit (Qiagen, Valencia, CA, USA). Real-time PCR analysis was performed for BRG1 binding sites in miR-148b promoter by using the following primers: 5′-AGC GCC AGT GTT AAA GGC TA-3′ (forward) and 5′-TCC ATG GGG AAC AGA AGA AG-3′ (reverse).
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