Protocols & Troubleshooting Guides

Immunohistochemistry, Western Blot, Enzyme Assays and other protocols

Apoptosis Assays

Caspase Activity Assays for Inhibitors of Apoptosis & Mitochondrial Proteins
Preparation of the Samples for Cytochrome c ELISA
Recombinant Human Bcl-2 Cytochrome c Release Assay
Recombinant Human Bcl-x
Recombinant Human Bcl-xL Cytochrome c Release
Recombinant Human BID Cytochrome c Release Assay
Recombinant Human BIM-L Cytochrome c Release Assay
Recombinant Human Caspase-8-cleaved BID Cytochrome c Release Assay
Recombinant Mouse Bcl-xL Cytochrome c Release Assay
Recombinant Mouse BID Cytochrome c Release Assay
Recombinant Mouse Caspase-8-cleaved BID Cytochrome c Release Assay

ELISA Protocols

ELISA Sample Preparation & Collection Guide
How to Adapt DuoSet ELISA Kits to 384-well Plate Format
How to Run an R&D Systems DuoSet ELISA
How to Run an R&D Systems Quantikine ELISA
How to Run an R&D Systems Quantikine™ QuicKit™ ELISA
Troubleshooting Guide: ELISA

Enzyme Assays

Enzyme Activity Assays using Fluorogenic Peptide Substrates
Inhibition of Calcineurin Phosphatase Activity
Recombinant Human ACE-2 Assay
Recombinant Human BMP-1/PCP Assay
Recombinant Human Carbohydrate Sulfotransferase 1/CHST1 Activity Assay
Recombinant Human Carbohydrate Sulfotransferase 2/CHST2 Activity Assay
Recombinant Human Carbohydrate Sulfotransferase 4/CHST4 Activity Assay
Recombinant Human Carbohydrate Sulfotransferase 6/CHST6 Activity Assay
Recombinant Human GalNAc4S-6ST/BRAG Activity Assay
Recombinant Human Meprin alpha Subunit/MEP1A Assay
Recombinant Human Sulfate 6-O-Sulfotransferase 1/HS6ST1 Activity Assay
Recombinant Mouse ACE-2 Assay
Recombinant Mouse Carbohydrate Sulfotransferase 1/CHST1 Activity Assay
Recombinant Mouse Carbohydrate Sulfotransferase 5/CHST5 Activity Assay
Recombinant Mouse Carbohydrate Sulfotransferase 7/CHST7 Activity Assay
Recombinant Mouse Heparan Sulfate 6-O-Sulfotransferase-3/HS6ST3 Activity Assay
Sulfotransferase Activity Assays

Flow Cytometry

7-Amino Actinomycin D (7-AAD) Cell Viability Flow Cytometry Protocol
Flow Cytometry Protocol for Cell Surface Markers
Intracellular Staining Flow Cytometry Protocol Using Alcohol Permeabilization
Intracellular Staining Flow Cytometry Protocol Using Detergents to Permeabilize Cells

ICC/IHC Protocols

Antigen Retrieval Methods
Appropriate Fixation of IHC/ICC Samples
Designing a Successful IHC/ICC Experiment
Detection & Visualization of Antibody Binding
Preparing Samples for IHC/ICC Experiments
Primary Antibody Selection & Optimization
Protocol for Heat-Induced Epitope Retrieval (HIER)
Protocol for Making a 4% Formaldehyde Solution in PBS
Protocol for the Preparation & Fixation of Cells on Coverslips
Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
Protocol for VisUCyte™ HRP Polymer Detection Reagent
The Importance of IHC/ICC Controls

Immunocytochemistry (ICC) Protocols

Protocol for the Preparation and Fluorescent ICC Staining of Cells on Coverslips
Protocol for the Preparation and Fluorescent ICC Staining of Non-adherent Cells
Protocol for the Preparation and Fluorescent ICC Staining of Stem Cells on Coverslips

Immunohistochemistry (IHC) Protocols

Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections
Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections
Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
Troubleshooting Guide: Immunohistochemistry

Immunology

CD4+ CD25+ Regulatory T Cell Isolation from PBMC or Splenocyte Preparations by Magnetic Selection
CD4+ T Cell Isolation from PBMC or Splenocyte Preparations by Magnetic Selection
CD4+ T Cell Isolation from PBMC or Splenocyte Preparations by T Cell Enrichment Column
Protocol for the Characterization of Human Regulatory T Cells
Protocol for the Characterization of Human Th22 Cells
Protocol for the Characterization of Human Th9 Cells
Protocol for the Differentiation and Characterization of Human Th1 Cells
Protocol for the Differentiation and Characterization of Human Th17 Cells
Protocol for the Differentiation and Characterization of Human Th2 Cells

Luminex

How to Run an R&D Systems Luminex® Assay
Luminex® Troubleshooting Guide

Neural Cell Culturing

Protocol for Culturing Embryonic Chick Dorsal Root Ganglion Neurons
Protocol for Culturing Embryonic Rat Spinal Motor Neurons
Protocol for Culturing Rat Cortical Neurons
Protocol for Culturing Rat Hippocampal Neurons
Protocol for Culturing Rat Microglia

Organoid and 3-D Cell Culture Protocols

Cultrex UltiMatrix BME Protocol
Culture of Mouse Enteric Organoids using Cultrex Basement Membrane Extract
Culture of Normal Human Gastric Organoids
Harvesting Organoids for Biochemical Analysis
Human Intestinal Organoid Culture Protocol
Human Liver Organoid Culture Protocol
Human Lung Organoid Culture Protocol
Immunofluorescence of Organoids Embedded in Cultrex Basement Membrane Extract
Use of Cultrex Basement Membrane Extract to Improve Take and Growth of Xenografts in Mice

Proteome Profiler Arrays

LI-COR Data Examples for Human Adipokine Array Kit
LI-COR Data Examples for Human Angiogenesis Array Kit
LI-COR Data Examples for Rat Cytokine Array Panel A
Profiling Changes in Receptor Tyrosine Kinase Phosphorylation using Antibody Arrays

Stem Cell Protocols

A Guide to the Colony Forming Cell Assay: Methods and Tips
Adipogenic Differentiation of Human/Mouse Mesenchymal Stem Cells
Chondrogenic Differentiation of Human/Mouse Mesenchymal Stem Cells
Culturing BG01V Human Embryonic Stem Cells on Irradiated Mouse Embryonic Fibroblasts (iMEF)
Culturing BG01V Human Embryonic Stem Cells with Mouse Embryonic Fibroblast (MEF)-Conditioned Media
Culturing Mouse Cortical Stem Cells: Expansion using the Neurosphere System
Culturing Rat Cortical Stem Cells: Expansion using the Monolayer System
Culturing Rat Cortical Stem Cells: Expansion using the Neurosphere System
Definitive Endoderm Differentiation of BG01V Human Embryonic Stem Cells
Endoderm, Ectoderm, and Mesoderm Differentiation of BG01V Human Embryonic Stem Cells
Mouse Hematopoietic Lineage Depletion
Osteogenic Differentiation of Human/Mouse Mesenchymal Stem Cells
Procedure for Serum-Free Expansion of Human Mesenchymal Stem Cells
Procedure for the Expansion of Human Mesenchymal Stem Cells
Protocol for the Preparation & Fixation of Stem Cells on Coverslips
Rat Cortical Stem Cell Culture Expansion Protocol
The Human Colony Forming Cell (CFC) Assay using Methylcellulose-based Media
The Mouse Colony Forming Cell (CFC) Assay using Methylcellulose-based Media
The Mouse Pre-B Colony Forming Cell (CFC) Assay Using Methylcellulose-based Media

Ubiquitin

Determine Ubiquitin Chain Linkage
Distinguish Between Poly-ubiquitination and Multi-mono-ubiquitination
Protocol for In Vitro Ubiquitin Conjugation Reaction

Video Protocols

Flow Cytometry Protocol for Staining Membrane Associated Proteins
How to Reconstitute Lyophilized Proteins
How to Run the CometAssay Analysis Software
Induction and Analysis of Epithelial to Mesenchymal Transition

Western Blot

R&D Systems Quality Control Western Blot Protocol
Troubleshooting Guide: Western Blot Figures
Western Blot Conditions
Western Blot Protocol for Cell Lysates
Western Blot Troubleshooting Guide