Testing NF-kappa B1-p50 antibody specificity using knockout mice. Journal Articles

abstract

• Cell extracts from knockout mice can provide definitive proof of antibody specificity. Two NF-kappa B1-p50 antibodies, sc-114 (a commercial antibody) and NR1157, were observed to recognize proteins having distinct electrophoretic mobilities of 52-55 kD and 50 kD, respectively, by Western blot analysis. In order to discriminate the specificity of these antibodies for NF-kappa B1-p50, whole cell extracts derived from NF-kappa B1-p105 knockout mice were employed. While the NR1157 antibody completely failed to recognize its 50 kD product in p105-/- knockout extracts, the sc-114 antibody still strongly recognized its 52-55 kD product. These data demonstrate that NR1157, but not sc-114, is highly specific for NF-kappa B1-p50 by Western blot analysis. In addition, these results highlight the utility of knockout cell extracts for discerning antibody specificity.

authors

• Pereira, DS
• Kushner, DB
• Ricciardi, RP
• Graham, Frank Lawson

status

• published 

publication date

• July 18, 1996

has subject area

• 1103 Clinical Sciences (FoR)
• 1112 Oncology and Carcinogenesis (FoR)
• Amino Acid Sequence (MeSH)
• Animals (MeSH)
• Antibodies (MeSH)
• Antibody Specificity (MeSH)
• Blotting, Western (MeSH)
• Cell Extracts (MeSH)
• Electrophoresis (MeSH)
• Humans (MeSH)
• Mice (MeSH)
• Mice, Knockout (MeSH)
• Molecular Sequence Data (MeSH)
• NF-kappa B (MeSH)
• NF-kappa B p50 Subunit (MeSH)
• Oncology & Carcinogenesis (Science Metrix)
• Rats (MeSH)

published in

• Oncogene  Journal

keywords

• Amino Acid Sequence
• Animals
• Antibodies
• Antibody Specificity
• Blotting, Western
• Cell Extracts
• Electrophoresis
• Humans
• Mice
• Mice, Knockout
• Molecular Sequence Data
• NF-kappa B
• NF-kappa B p50 Subunit
• Rats

PubMed ID

• 8710386

start page

• 445

end page

• 446

volume

• 13

issue

• 2