Tissues were isolated and fixed using 4% paraformaldehyde and then embedded in paraffin. Antigen retrieval was performed using the 2100 Antigen Retriever in buffer A (Electron Microscopy Sciences), and standard immunostaining procedures were performed for Dnmt1 (Santa Cruz Biotechnology), Chga (Immunostar), Dnmt3a (Santa Cruz Biotechnology), Dnmt3b (Abcam), E-Cadherin (BD Transduction Laboratories), Sox9 (Millipore), Lyz (Dako), Ki67 (BD Pharmingen), and Muc2 (Santa Cruz Biotechnology). Immunohistochemical procedures were modified for Hes-1 (Ben Stanger, University of Pennsylvania), including antigen retrieval in high-pH antigen unmasking solution (Vector Laboratories) and signal amplification with the TSA Fluorescein system (Perkin Elmer). AP staining was performed using NBT and BCIP (Boehringer Ingelheim). In situ hybridization for Olfm4 was performed as described previously (Barker et al. 2007 (link)). All microscopy was performed on the Nikon Eclipse 80i.
2100 antigen retriever in buffer a
Manufactured by Electron Microscopy Sciences
The 2100 Antigen-Retriever in Buffer A is a laboratory equipment designed for the pretreatment of tissue samples prior to immunohistochemical (IHC) analysis. It provides a controlled heating process to facilitate the retrieval of antigenic sites, which is a crucial step in IHC protocols. The equipment operates using a specific buffer solution, Buffer A, to optimize the antigen retrieval process.
Automatically generated - may contain errors
Lab products found in correlation
Dnmt1, by Santa Cruz Biotechnology (3 mentions)
E cadherin, by BD (3 mentions)
Dnmt3a, by Santa Cruz Biotechnology (2 mentions)
γh2ax, by Cell Signaling Technology (2 mentions)
Tunel label and enzyme, by Roche (2 mentions)
Dnmt3b, by Abcam (1 mentions)
Tsa fluorescein system, by PerkinElmer (1 mentions)
High ph antigen un masking solution, by Vector Laboratories (1 mentions)
Dnmt3b, by Novus Biologicals (1 mentions)
Epcam, by Abcam (1 mentions)
β catenin, by BD (1 mentions)
Eclipse 80i, by Nikon (1 mentions)
3 protocols using 2100 antigen retriever in buffer a
1
Comprehensive Immunohistochemical Analysis of Intestinal Tissue
2
Immunohistochemical Analysis of Epigenetic Markers
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Tissues were isolated and fixed using 4% paraformaldehyde in PBS and then embedded in paraffin. Antigen retrieval was performed using the 2100 Antigen-Retriever in Buffer A (Electron Microscopy Sciences, Hatfield, PA) and standard immunostaining procedures were performed for Dnmt1 (Santa Cruz), Dnmt3a (Santa Cruz Biotechnology, Dallas, TX), Dnmt3b (Imgenex, San Diego, CA), E-Cadherin (BD Biosciences, San Jose, CA), Ki67 (BD Biosciences), and γH2AX (Cell Signaling Technology, Berverly, MA). TUNEL staining was performed using TUNEL Label and Enzyme (Roche, Indianapolis, IN) and AlexaFluor 555-aha-dUTP (Molecular Probes, Eugene, OR). All microscopy was performed on a Nikon Eclipse 80i (Tokyo, Japan). For all immunofluorescence and immunohistochemistry staining, n=3 biological replicates per genotype, per timepoint.
3
Immunostaining and TUNEL Analysis
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