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Human, reacts against Thr76/Thr84-phosphorylated nucleolin
Immunogen:
Modified peptide
Formulation:
This antibody is provided in phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and 50% glycerol. Final antibody concentration is 0.5 mg/ml.
Each lot of this antibody is quality control tested by Western blotting. Western blotting, suggested working dilution(s): Use 10 μl per 5 ml antibody dilution buffer for each mini-gel. For IHC, use 10 μg/ml of antibody for staining. Antigen retrieval using 0.01 M sodium citrate buffer is recommended. It is recommended that the reagent be titrated for optimal performance for each application.
COA:
Application References:
1. Tediose T, et al. 2010. Nucleic Acids Res. 38:2799. PubMed 2. Kolev MV, et al. 2010. J. Immunol. 184:6035. PubMed
Formalin-fixed paraffin-embedded human testis tissue was stained with 10C7 at 10 µg/ml and developed with an alkaline phosphatase chromogen substrate (red color). Tissue was counterstained with H&E (blue/pink). Magnification, 40X.
Hela cell nuclear extract (2 x 106 cells) was phosphorylated in vitro using cdc2 kinase and ATP for 10 min. at 30C. Lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with anti-phospho-nucleolin (Clone 10C7) antibody. Proteins were visualized using a goat anti-mouse secondary conjugated to HRP and a chemiluminescence detection system. Lane 1 (-), no cdc2-kinase/ATP addition; Lane 2 (+), cdc2 kinase/ATP added.
Nucleolin is a ubiquitously expressed 100 kD nuclear and nucleolar protein that contains RNA recognition motifs. Nucleoloin is the major nucleolar protein of eukaryotic cells in growth phase associated with intranucleolar chromatin and pre-ribosomal particles. This protein induces chromatin de-condensation by binding to histone H1 and plays a role in pre-rRNA transcription and ribosome assembly. Nucleolin exhibits self-cleaving, DNA helicase, RNA helicase and DNA-dependent ATPase activities; activity can be upregulated in response to ERK signaling, c-Myc expression, and genotoxic stress (UV radiation). This protein is modified by phosphorylation (Thr76/Thr84). This protein interacts with phosphatase 1δ, hepatitis C virus NS5B, part of B-cell specific transcription factor complex with LR1, and DNA Topoisomerase I. The 10C7 monoclonal antibody recognizes human phosphorylated nucleolin (Thr76/Thr84) and has been shown to be useful for Western blotting.
Other Names:
Nucleolin protein, C23
Structure:
RNA recognition motifs; 100 kD
Distribution:
Nuclear, nucleolar; ubiquitously expressed
Function:
Major nucleolar protein of eukaryotic cells in growth phase associated with intranucleolar chromatin and pre-ribosomal particles. Induces chromatin decondensation by binding to histone H1. Plays a role in pre-rRNA transcription and ribosome assembly; exhi
Regulation:
Upregulated in response to ERK signaling, c-Myc expression, genotoxic stress (UV radiation)
Modification:
Phosphorylation
Interaction:
Protein phosphatase 1δ, hepatitis C virus NS5B, part of B-cell specific transcription factor complex with LR1, interacts with DNA Topo I
Antigen References:
1. Srivastava M, et al. 1989. FEBS Lett. 250:99. 2. Westmark C, et al. 2001. J. Biol. Chem. 276:1119. 3. Ying G, et al. 2000. J. Biol. Chem. 275:4152. 4. Hanakahi L, et al. 1997. P. Natl. Acad. Sci. USA 94:3605.
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Formalin-fixed paraffin-embedded human testis tissue was stained with 10C7 at 10 µg/ml and developed with an alkaline phosphatase chromogen substrate (red color). Tissue was counterstained with H&E (blue/pink). Magnification, 40X.
Hela cell nuclear extract (2 x 106 cells) was phosphorylated in vitro using cdc2 kinase and ATP for 10 min. at 30C. Lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with anti-phospho-nucleolin (Clone 10C7) antibody. Proteins were visualized using a goat anti-mouse secondary conjugated to HRP and a chemiluminescence detection system. Lane 1 (-), no cdc2-kinase/ATP addition; Lane 2 (+), cdc2 kinase/ATP added.
