Recommended Antibody Protocols

alamarBlue Protocols

Detailed guides to help you use alamarBlue

• QC test for alamarBlue
• Method to determine LD50 using alamarBlue
• General method for measuring cytotoxicity or proliferation using alamarBlue (spectrophotometry method and fluorescence method)
• Method for determining optimum length of incubation and plating density
• Spectrophotometry calculations for using different filters
• alamarBlue example experiments

MCM2 Protocol

Detailed guide to help you use MCM2

• Rabbit anti-MCM2 proliferation pairs staining procedure

ELISA Protocols

General ELISA protocols

• Indirect
• Competition (Inhibition)
• Direct ELISA with streptavidin-biotin detection
• Sandwich ELISA with streptavidin-biotin detection
• Sandwich ELISA with direct detection

Protocols for use with anti-biotherapeutic antibodies

• PK bridging ELISA measuring free drug
• PK bridging ELISA measuring total drug
• PK antigen capture ELISA measuring bound drug exclusively
• ADA bridging ELISA

Flow Cytometry (FC) Protocols

• Preparation of tissue culture cell lines for flow cytometry
• Preparation of peripheral blood mononuclear cells for flow cytometry
• Preparation of peritoneal macrophages, bone marrow, thymus, and spleen cells for flow cytometry
• Direct immunofluorescence staining of surface epitopes of cells and whole blood
• Indirect immunofluorescence staining of surface epitopes of cells and whole blood
• Direct immunofluorescence staining of immunoglobulin light chains on B lymphocytes in whole blood
• Direct immunofluorescence staining of intracellular antigens: the Leucoperm method
• Direct immunofluorescence staining of intracellular antigens: the methanol plus Leucoperm method
• Direct immunofluorescence staining of intracellular cytokines in whole blood
• Direct immunofluorescence staining of intracellular antigens: the digitonin method
• Direct immunofluorescence staining of intracellular antigens: the paraformaldehyde/saponin method
• Propidium iodide staining of cells for cell cycle analysis
• Propidium iodide staining of cells for cell cycle analysis in combination with direct immunofluorescence intracellular staining
• BrdU staining of cells for cell cycle analysis
• Unprimed T-cell activation: pharmacologic methods
• Unprimed T-cell activation: antibody stimulation method
• Primed T-cell activation: antigen-presenting cell co-culture method
• Measuring cell proliferation using CytoTack, a cell-permeable dye
• Direct immunofluorescence staining of intracellular antigens using antibodies labeled with StarBright™ Dyes
• Direct immunofluorescence staining of whole blood for intracellular CD68: the Leucoperm method
• Direct immunofluorescence staining of cells with StarBright Dye-conjugated antibodies

Immunofluorescence (IF) Protocols

• Fluorescence microscopy (Direct immunofluorescence) for cell suspensions
• BrdU Labeling of heLa cells followed by immunostaining with Mouse Anti-BrdU Antibody
• BrdU labeling of heLa cells followed by immunostaining with Rabbit Anti-BrdU Antibody

Immunohistochemistry (IHC) Protocols

Protocols for IHC-frozen and IHC-paraffin

• Indirect Immunostaining of frozen tissue sections 
• Streptavidin-biotin immunostaining of frozen tissue sections 
• PAP/APAAP Immunostaining of frozen tissue sections 
• Indirect immunostaining of paraffin-embedded tissue sections 
• Streptavidin-biotin immunostaining of paraffin-embedded tissue sections 
• PAP/APAAP immunostaining of paraffin embedded tissue sections 
• Antigen retrieval techniques for use with formalin-fixed paraffin-embedded sections 
• Antigen retrieval techniques: For use with F4/80 immunostaining of paraffin-embedded sections 
• Fixative Recipes

Western Blot (WB) Protocols

• Western Blotting protocol 
• Detection of phosphorylated proteins by Western Blotting
• Method to avoid interference from denatured IgG using TidyBlot Western Blot detection reagent
• Fluorescent Western Blotting Protocol Using StarBright Dye Secondary Antibodies