Protocols and troubleshooting tips

Protocols - by application

Antibody conjugation (1)+

• Biotin conjugation

Cell culture (4)+

• Cell counts using a haemocytometer
• Cell culture guidelines
• Dealing with cell culture contamination
• Freezing cells

Cell isolation (1)+

• T cell purification from splenocytes

ChIP (7)+

• ChIP tips
• ChIP troubleshooting tips
• ChIP using plant samples - Arabidopsis
• Chromatin preparation from tissues for chromatin immunoprecipitation (ChIP)
• Cross-linking chromatin immunoprecipitation (X-ChIP) protocol
• Native chromatin immunoprecipitation protocol
• X-ChIP protocol summary

Dot blot (1)+

• Dot blot protocol

ELISA (9)+

• Calculation of ELISA results from the standard curve.
• Control samples required for ELISA
• Direct ELISA using fluorescent substrate protocol
• Direct ELISA using primary antibody protocol
• ELISA troubleshooting tips
• Indirect ELISA protocol
• Indirect ELISA protocol summary
• Sandwich ELISA protocol
• Sandwich ELISA protocol summary

ELISPOT (3)+

• Dual ELISPOT
• ELISPOT protocol
• ELISPOT troubleshooting tips

Flow cytometry / FACS (11)+

• Direct flow cytometry (FACS) protocol
• Flow cytometry - recommended controls
• Flow cytometry immunophenotyping
• Flow cytometry intracellular staining protocol
• Flow cytometry membrane staining protocol summary
• Flow cytometry troubleshooting tips
• Fluorescence activated cell sorting of live cells
• Indirect flow cytometry (FACS) protocol
• Intracellular flow cytometry protocol summary
• Introduction to flow cytometry
• Red blood cell lysis protocol

Immunoprecipitation (4)+

• Crosslinking the antibody to the beads protocol
• Immunoprecipitation protocol
• IP troubleshooting tips
• Using IgM antibodies for IP

Immunostaining (18)+

• Determining if antibody binds only phosphorylated protein
• Double immunofluorescence: sequential protocol
• Double immunofluorescence: simultaneous protocol
• Fixation and permeabilization for IHC and ICC
• GMA embedding for immmunohistochemistry
• IHC frozen sections staining protocol
• IHC paraffin staining protocol
• IHC Protocol Summary
• IHC troubleshooting tips
• Immunocytochemistry and immuofluorescence protocol
• Immunohistochemistry - Methods Express Book Series
• In situ hybridization
• Mouse on mouse staining procedure
• Perfusion fixation protocol
• Perfusion fixation troubleshooting tips
• Preparing cells and sections for BrdU immunostaining
• Recommended controls for IHC/ICC
• Whole mount staining protocols

siRNA  (1)+

• Recommended checks and controls for siRNA experiments

Toxin neutralization (2)+

• Cell preparation/plating protocol
• Toxin neutralization assay

Western blot (15)+

• Western blotting - a detailed guide

• Blocking with immunizing peptide protocol
• Buffers and stock solutions
• De-phosphorylation of proteins protocol
• Histone blotting protocol
• Histone extraction protocol
• Mitochondrial purification protocol
• Nuclear fractionation protocol
• Peptide competition assay (peptide blocking)
• Phospho-proteins protocol
• Recommended controls required for WB
• Soluble (S-100) mitochondrial fractionation protocol
• Stripping for reprobing protocol
• Western blot protocol summary
• Western blot troubleshooting tips

Protocols - by research area

Apoptosis protocols (5)+

• Annexin V detection protocol
• Apoptosis induction protocols
• Caspase detection protocol
• DNA fragmentation analysis protocol
• Introduction to apoptosis

Cardiovascular protocols (2)+

• Cardiomyocyte immunofluorescence protocol
• Mouse adipocyte glucose uptake assay

Molecular biology protocols (4)+

• Calcium phosphate transfection protocol
• RNA isolation protocol: cells in culture
• Subcellular fractionation protocol
• Transforming competent cells and isolating plasmid DNA

Neuroscience protocols (11)+

• Amyloid Beta Uptake by Glia Cells
• Antibody staining of whole mount Drosophila embryos
• Bielschowsky’s Silver Stain for Axons
• Brain Synaptosomal Preparation
• BrdU protocol
• DEA Extraction of [soluble] Amyloid beta from Mouse Brain
• FA Extraction of [insoluble] Amyloid beta from Mouse Brain
• IF protocol for perfusion fixed tissue
• IHC Protocol for Free Floating Brain Sections
• Nuclear enrichment
• Primary Cultures for IHC – Viability Assays

Protocol guides and diagrams

Detailed application guides (3)+

• Cell culture guidelines
• IHC paraffin staining guide
• Western blotting guide

Protocol diagrams (8)+

• ELISPOT protocol summary
• Flow cytometry membrane staining protocol summary
• IHC protocol summary
• Indirect ELISA protocol summary
• Intracellular flow cytometry protocol summary
• Sandwich ELISA protocol summary
• Western blot protocol summary
• X-ChIP protocol summary

General help

Frequently asked questions (6)+

• ChIP FAQ
• ELISA FAQ
• Flow cytometry FAQ
• Immunohistochemistry FAQ
• Immunoprecipitation FAQ
• Western blot FAQ

Product guides (4)+

• ELISA, ELIPAIR, ELISPOT & FLUOROSPOT Kits
• Immunohistochemistry
• Proteins and enzymes
• Secondary antibodies

Secondary antibody and fluorescence guides (6)+

• DyLight® conjugated secondary antibodies
• DyLight® fluorochromes
• Fluorescence guide
• Fluorophore table
• How to choose a conjugate
• How to choose the right secondary antibody

Troubleshooting tips (7)+

• ChIP troubleshooting tips
• ELISA troubleshooting tips
• ELISPOT troubleshooting tips
• Flow cytometry troubleshooting tips
• IHC troubleshooting tips
• IP troubleshooting tips
• Western blot troubleshooting tips