Nucleolin Antibody for Flow Cytometry / NCL Antibody (FACS) [clone NCL/902] (V2755)

Description

Nucleolin (NCL) is a multifunctional nucleolar phosphoprotein that plays a central role in ribosome biogenesis, ribosomal RNA transcription, and nucleolar organization. Encoded by the NCL gene on chromosome 2q37, nucleolin is one of the most abundant proteins within the nucleolus of proliferating cells and participates in ribosomal RNA synthesis, RNA processing, and ribonucleoprotein complex assembly. The Nucleolin Antibody for Flow Cytometry clone NCL/902 is a mouse monoclonal reagent developed for analysis of nucleolin expression using flow cytometry and fluorescence-activated cell sorting (FACS) assays.

Flow cytometry enables rapid quantitative analysis of protein expression across large populations of individual cells. When used as an NCL Antibody (FACS), nucleolin antibodies allow researchers to measure intracellular NCL levels, identify proliferating cell populations, and evaluate changes in nucleolar protein expression during cellular activation or transformation. Because nucleolin is primarily localized within the nucleus and nucleolus, detection by flow cytometry typically involves fixation and permeabilization steps that allow antibodies to access intracellular proteins prior to FACS analysis. Fluorescently labeled antibodies or fluorophore-conjugated secondary reagents then allow nucleolin-positive cells to be detected and quantified by fluorescence intensity during flow cytometric analysis.

Because nucleolin is highly enriched within nucleoli and displays a characteristic nuclear localization pattern, antibodies targeting NCL are also widely used in complementary experimental approaches including immunofluorescence microscopy, immunohistochemistry staining of tissue sections, and western blot analysis of nucleolin expression. Immunofluorescence can reveal the punctate nucleolar distribution of nucleolin within cells, while immunohistochemistry allows visualization of nuclear nucleolin expression in formalin-fixed tissue samples. Western blot experiments further support detection of nucleolin protein levels in cell lysates. Incorporation of these complementary techniques alongside flow cytometry provides a broader understanding of nucleolin regulation across different biological systems.

Nucleolin contains several structural domains that enable its diverse biological functions. The N-terminal acidic region interacts with chromatin and ribosomal RNA transcription complexes. Four central RNA recognition motifs allow nucleolin to bind ribosomal RNA and other RNA molecules, while the glycine- and arginine-rich C-terminal region mediates interactions with nucleic acids and nucleolar proteins. Through these domains nucleolin coordinates ribosomal RNA synthesis, processing, and ribonucleoprotein assembly, processes essential for ribosome production and cellular growth.

NCL antibody reagents are commonly described in the literature using several established synonyms for the protein. NCL antibody, also known as nucleolin antibody, C23 nucleolin antibody, and nucleolar protein nucleolin antibody, recognizes a nucleolar phosphoprotein historically referred to as nucleolin nucleolar phosphoprotein. The designation C23 originates from early nucleolar protein fractionation studies that identified nucleolin as a prominent nucleolar phosphoprotein associated with ribosomal transcription complexes. These alternative names remain widely used in research focused on nucleolar biology and ribosome synthesis.

Nucleolin expression is closely associated with cellular proliferation and nucleolar activity. Elevated levels of NCL are frequently observed in rapidly dividing cells and many tumor types where increased ribosome production supports enhanced protein synthesis. Because flow cytometry allows quantitative analysis of fluorescence intensity in individual cells, a Nucleolin Antibody for Flow Cytometry such as clone NCL/902 provides a valuable tool for examining nucleolin expression patterns, analyzing heterogeneous cell populations, and studying nucleolar protein regulation in cancer biology and cell growth studies.

Application Notes

Optimal dilution of the Nucleolin antibody should be determined by the researcher.

1. Staining of formalin-fixed tissues requires boiling tissue sections in pH 9 10mM Tris with 1mM EDTA for 10-20 min followed by cooling at RT for 20 min
2. The prediluted format is supplied in a dropper bottle and is optimized for use in IHC. After epitope retrieval step (if required), drip mAb solution onto the tissue section and incubate at RT for 30 min.

Immunogen

Recombinant human protein was used as the immunogen for the Nucleolin antibody.

Storage

Store the Nucleolin antibody at 2-8oC (with azide) or aliquot and store at -20oC or colder (without azide).

Alternate Names

NCL antibody, nucleolin antibody, C23 nucleolin antibody, nucleolar protein nucleolin antibody, nucleolin nucleolar phosphoprotein antibody