Fundamental Techniques

Primarily for use in teaching and as a vehicle for students to demonstrate the quality of their work, this type of protocol usually covers established laboratory techniques.

Staining for γH2AX in paraffin-embedded tissue sections

Staining for γH2AX in paraffin-embedded tissue sections

Staining for γH2AX in paraffin-embedded tissue sections This protocol describes how to stain sections of formalin-fixed paraffin-embedded tissue for phosphorylated Ser139 on histone variant H2A.X (γH2AX). A primary antibody is used to identify and bind to γH2AX, and a secondary antibody conjugated to a fluorescent probe identifies and binds the primary antibody for imaging by […]

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Photolithography

Photolithography

Photolithography Engelward Lab protocol for creating microfeature molds from silicon wafers by lithographically patterning SU-8 photoresist.  

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Staining for Ki67 in paraffin-embedded tissue sections

Staining for Ki67 in paraffin-embedded tissue sections

Staining for Ki67 in paraffin-embedded tissue sections This protocol describes how to immunofluorescence stain for Ki67, a marker of cellular proliferation, in paraffin-embedded tissue sections.  

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PCR to genotype RaDR mouse from ear punch

PCR to genotype RaDR mouse from ear punch

PCR to genotype RaDR mouse from ear punch These protocols describe the process to genotype for the RaDR transgene in mouse ear punches. RaDR mice were developed in the Engelward lab at MIT, and they allow detection of sequence rearrangement mutations arising from aberrant homologous recombination in situ. Please see the paper below for more […]

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Disaggregation of intact colon crypts

Disaggregation of intact colon crypts

Disaggregation of intact colon crypts This protocol describes how to disaggregate whole colon tissue into a suspension of colonic crypts. This protocol was adapted from Samuel et al., Am J Physiol Cell Physiol 296: C296-C305, 2009. The sample image shows a suspension of crypts, with intact crypts outlined in red.  

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Western Blot with BioRad Protean 3 Cell

Western Blot with BioRad Protean 3 Cell

Western Blot with BioRad Protean 3 Cell This protocol describes how to perform a Western blot using the BioRad Protean 3 Cell system, one of many available Western blot assemblies. The basic steps of this protocol can be adapted to any Western blot assembly, but we recommend reading the instructions for your system first.  

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Immunofluorescent staining on paraffin-embedded tissue sections

Immunofluorescent staining on paraffin-embedded tissue sections

Immunofluorescent staining on paraffin-embedded tissue sections This protocol describes the steps necessary to fluorescently detect proteins in paraffin-embedded tissue sections. After depariffinizing and rehydrating the tissues, antigens are exposed through incubation with boiling antigen retrieval buffer. Subsequently, sections are blocked with bovine serum albumin and probed with primary antibody overnight. After a series of washes, […]

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Whole blood hemolysis with isotonic ammonium chloride solution

Whole blood hemolysis with isotonic ammonium chloride solution

Whole blood hemolysis with isotonic ammonium chloride solution  

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