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Protocols and troubleshooting tips

Protocols and troubleshooting tips on how to get the best from our antibodies

  • Antibody storage guide
  • Need help or want to give feedback on our protocols? Contact Abcam's Scientific Support Team

Protocols - by application

Antibody conjugation and purification (2)+
  • Sodium Azide removal protocol

  • Biotin conjugation
Cell culture (7)+
  • Aseptic technique video protocol
  • Cell culturing summary
  • Counting cells using a haemocytometer
  • Cryopreservation of mammalian cell lines
  • Differentiation of Human Embryonic Stem Cells to Hepatic Endoderm
  • Mammalian cell tissue culture techniques
  • Stimulating MMP-9 expression protocol
Cell isolation (1)+
  • Murine endothelial cell isolation

ChIP (7)+
  • Discussion of ChIP protocol and technique

  • 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 (10)+
  • Calculating & evaluating ELISA data
  • Control samples required for ELISA
  • ELISA sample preparation guide
  • ELISA troubleshooting tips
  • In Cell ELISA (ICE)
  • Indirect ELISA protocol
  • Indirect ELISA protocol summary
  • Non-denaturing extraction for ELISA and dipstick activity assays
  • Sandwich ELISA protocol
  • Sandwich ELISA protocol summary
ELISPOT (2)+
  • Dual ELISPOT protocol: Detection of two secreted targets per sample
  • ELISPOT protocol
Flow cytometry / FACS (13)+
  • 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
  • Fluorescence compensation in flow cytometry
  • Indirect flow cytometry (FACS) protocol
  • Intracellular flow cytometry protocol summary
  • Introduction to flow cytometry
  • Propidium iodide staining of cells to assess DNA cell cycle
  • Red blood cell lysis protocol
Immunoprecipitation (6)+
  • DNA-RNA immunoprecipitation (DRIP) protocol
  • Immunoprecipitation protocol
  • IP troubleshooting tips
  • RNA Immunoprecipitation (RIP)
  • Using IgM antibodies for IP
  • UV Cross-Linking and Immunoprecipitation (CLIP)
Immunostaining (18)+
  • BrdU immunostaining procedure for cell cultures and tissue sections.

  • GMA (Glycol methacrylate) embedding for immmunohistochemistry protocol

  • ICC/IF Immunocytochemistry and immuofluorescence protocol

  • IHC and ICC fixation and permeabilization protocol 

  • IHC-FoFr Immunohistochemistry perfusion fixation protocol

  • IHC-FoFr Perfusion fixation troubleshooting tips

  • IHC-Fr frozen sections staining protocol

  • IHC-FrFl Immunohistochemistry on free floating brain sections

  • IHC-P Immunohistochemistry protocol

  • ISH - In situ hybridization protocol

  • Mouse on Mouse (MOM) staining procedure

  • Validating phospho-specificity of antibodies (WB, IHC) protocol

  • IHC for paraffin-embedded sections video protocol
  • IHC troubleshooting tips
  • Immunohistochemistry - Methods Express Book Series
  • Recommended controls for IHC/ICC
  • Sectioning of paraffin-embedded tissue video protocol
  • Whole mount staining protocols
siRNA  (1)+
  • Recommended checks and controls for siRNA experiments
Toxin neutralization (2)+
  • Cell culture preparation and plating protocol

  • Toxin neutralization assay
Western blot (17)+
  • Western blotting - a detailed guide

  • Blocking with immunizing peptide protocol
  • Blue native electrophoresis protocol
  • Buffers and stock solutions
  • Histone blotting protocol
  • Histone extraction protocol for Western blot
  • Mitochondrial purification protocol for Western blot samples
  • Peptide competition assay (peptide blocking)
  • Procedure for detection of phosphorylated proteins in Western blot
  • Recommended controls for Western blotting
  • Sample preparation for western blot
  • Soluble (S-100) mitochondrial fractionation protocol for Western blot
  • Western blot membrane stripping for restaining protocol
  • Western blot protocol
  • Western Blot protocol for high molecular weight proteins (150 - 300 kDa)
  • Western blot short step by step diagram
  • Western blot troubleshooting tips

Protocols - by research area

Apoptosis protocols (5)+
  • Annexin V detection protocol for apoptosis
  • Apoptosis DNA fragmentation analysis protocol
  • Caspase apoptosis detection protocol
  • Introduction to apoptosis
  • Protocols for biological and chemical induction of apoptosis in cells
Cardiovascular protocols (3)+
  • Cardiomyocyte immunofluorescence protocol
  • Isolation of human platelets from whole blood
  • Mouse adipocyte glucose uptake assay
Molecular biology protocols (5)+
  • Gelatin zymography protocol
  • Measuring activity of gelatinase enzymes
  • RNA isolation protocol: cells in culture
  • Subcellular fractionation protocol
  • Transforming competent cells and isolating plasmid DNA
Neuroscience protocols (12)+
  • Amyloid Beta Uptake by Glia Cells
  • Antibody staining of whole mount Drosophila embryos
  • Bielschowsky’s Silver Stain for Axons
  • BrdU protocol
  • CLARITY 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
  • SWITCH protocol
Stem cells protocols (3)+
  • Differentiation of 3T3-L1 cells into adipocyte-like cells
  • Hepatocyte differentiation protocol
  • Mouse embryonic stem cells to macrophage-like cells

Protocol guides and diagrams

Detailed application guides (4)+
  • MitoTox™ : Mitochondrial toxicity application guide

  • 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

Product guides (5)+
  • Antibody arrays - specific protein profiling tool

  • ELISA, ELIPAIR, ELISPOT & FLUOROSPOT Kits
  • Fluorescent ligand binding - frequently asked questions
  • Immunohistochemistry (IHC) reagents | IHC protocols
  • Secondary antibodies
Secondary antibody and fluorescence guides (6)+
  • Conjugate guide | Biotin, HRP, AP and Fluorochrome
  • DyLight® Fluorochrome conjugated secondary antibodies
  • Fluorescence guide
  • Fluorophore table
  • The complete guide to secondary antibody selection
  • Understanding secondary antibodies
Troubleshooting tips (7)+
  • Troubleshooting tips for enzymatic assay kits

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

Other useful resources

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