Introduction to Immunocytochemistry (House Ear Institute)
A brief overview of common available methods.
BrDU Immunocytochemistry using peroxidase and DAB for rat tissues (Contributed by Shyam Laxminarayan)
BrDU (5-Bromo-2-Deoxy uridine) is a synthetic Thymidine analog which incorporates into single stranded DNA during the S-Phase of the Cell Cycle. Antibodies against BrDU can be used along with a detection system (such as Peroxidase/DAB) to immunostain for BrDU in mammary gland sections to reveal the distribution of actively dividing cells, hence the % of proliferating cells.
Estrogen Receptor/Progesterone Receptor Immunocytochemistry (Contributed by Shyam Laxminarayan)
Protocol specifically for detecting estrogen/progesterone receptor by immunocytochemistry.
Immunocytohistochemistry (Howell Lab)
Procedures for cell plating, fixing, antibody incubation and mounting
· Immunocytochemical Identification of Proliferating Cells in Vascular Tissue (anti-BrdU); Vector Red Detection(Wilcox Lab)
· Immunocytocheimical Identidication of Proliferating Cells in Vascular Tissue (anti-BrdU); Diaminobenzadine (DAB) Brown Detection (Wilcox Lab)
Immunohisto- and Immunocyto-chemisty-Enzymatic Protocol(R&D)
General and nice protocol for immunocyto and immunohistochemistry including sample preparation, staining and trouble shooting
Immunocytochemisty-Fluorescence Protocol (R&D)
General protocol including details on staining cell in suspension and on slide
Embedding in Resins for Immunocytochemistry (House Ear Institute)
Describes the use of the specialized and not so specialized resins.
Freeze Substitution for Embedding in Resins (House Ear Institute)
A simple, inexpensive way of embedding in resin using freeze substitution methods.
Using Resin Sections for Immunofluorescence (House Ear Institute)
A protocol for applying antibodies with fluorescent markers onto sections of resin-embedded material. The thin sections help increase resolution.
Preparation of Colloidal Gold Probes (House Ear Institute)
See how easily, and inexpensively, these expensive probes can be prepared. Excellent information if large volumes of gold probes are required.
Silver Enhancement of Colloidal Gold (House Ear Institute)
How to precipitate silver onto small colloidal gold particles
so that the signal can be detected by light microscopy.
Problems with Immunolabeling (House Ear Institute)
When labeling cells or sections with antibodies or other affinity markers, the results are not easily interpreted. Look here for some help.
The Use of Affinity Cytochemistry to Detect Ligands Bound to Target Cells (Childs Lab)
This protocol is used to detect neuropeptides and growth factors bound to target sites on pituitary cells. Pituitary cells are grown in culture for 1-5 days (time dependent on the experimental design). The cells are exposed to physiological concentrations of biotinylated ligands for 1 min to 4 h, depending on the type of experiment. After exposure, the cells are fixed for light or electron microscopic detection of the site of binding.
Preparing coated coverslips (Hoshi Lab)
Some cells stick to coveslips better if they are coated with poly amino acids or collagen. Poly-(D, L, D/L)-lysine is commonly used to treat cover slips.