Table 3.

Dilution guidelines for unpurified antibody preparations

Unpurified antibodies will not have a concentration stated on the vendor datasheet. For most whole antisera, culture supernatants, or ascites fluid products, the concentration may be unknown. Unpurified antibody preparations vary significantly in specific antibody concentrations. If the specific antibody concentration of an unpurified antibody preparation is unknown, these concentration estimates may be used as a rough guideline. Please remember that these dilutions and concentration estimates are only a starting point, and dilutions may need to be adjusted based on the experimental results. IHC, immunohistochemistry; ICC, immunocytochemistry; EIA, enzyme immunoassay. NA, not applicable.

Assay	Tissue culture supernatant	Ascites	Whole antiserum	Purified antibody
Western blot/dot blot	1:100	1:1000	1:500	1 μg/ml
IHC/ICC	Neat: 1:10	1:100	1:50–1:100	5 μg/ml
EIA/ELISA	1:1000	1:10,000	1:500	0.1 μg/ml
FACS/flow cytometry	1:100	1:1000	1:500	1 μg/ml
Immunoprecipitation	NA	1:100	1:50–1:100	1–10 μg/ml
Concentration estimate	1–3 mg/ml	5–10 mg/ml	1–10 mg/ml	NA