Nuclear Marker Antibodies
The nucleus is clearly apparent in all eukaryotic cells. It is bound by a double membrane and contains most of the cell’s genetic material, which consists of DNA molecules complexed with proteins to form chromosomes. The nucleus functions to maintain the genes within these chromosomes and to regulate their expression, thereby controlling the activities of the cell.
Nuclear pores provide a channel across the otherwise impermeable nuclear membrane, through which small molecules and ions can freely move, and large molecules can be actively transported by carrier proteins. While the nucleus does not contain any membrane bound sub compartments, it does include a number of unique bodies, the nucleolus being the most easily identified.
The nucleolus is the site of ribosomal RNA transcription and ribosome assembly. Nuclear organizing regions (NORs), which are made up of tandem repeats of rRNA genes, form the foundation of the nucleolar structure.
Chromatin, which consists of DNA, RNA, and protein, is also found in the nucleus and is organized into chromosomes. Centromeres serve to link chromosome pairs, and the centromere is the site of spindle fiber attachment via kinetochores during mitosis.
Nuclear marker antibodies detect proteins specific to the nucleus and can aid in the study of the morphology and dynamics of the nucleus and its structures. Nuclear marker antibodies also provide a way for monitoring nuclear changes throughout cellular processes.
Each Invitrogen nuclear marker antibody is validated for use in applications such as ELISA, western blot, flow cytometry, immunofluorescence, immunohistochemistry, immunocytochemistry, and immunoprecipitation.
Nuclear marker antibody targets
Featured product data
Immunofluorescent analysis of HIF-1 alpha using HIF-1 alpha Monoclonal Antibody (mgc3) (Cat. No. MA1-516) shows staining in U251 cells. HIF-1 alpha (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with an antibody recognizing HIF-1 alpha (Cat. No. MA1-516) at a dilution of 1:20 overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Cat. No. 35552 for GAR, Cat. No. 35503 for GAM). Images were taken at 60X magnification.
Immunohistochemistry was performed on normal deparaffinized human heart tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH 6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval, tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a rabbit polyclonal antibody recognizing iNOS (Cat. No. PA1-036) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
Sandwich ELISA analysis of human Interferon Gamma (IFNG) was performed using a Human Interferon Gamma Colorimetric ELISA kit by loading 50 ul per well of Interferon Gamma Recombinant Protein (Cat. No. RIFNG100) in quadruplicate at 1000, 400, 160, 64, 25.6, and 0 pg/ml across a 3 ug/ml mouse anti-human IFNG (Cat. No. M700A) pre-coated plate and incubating for 2 hours at room temperature. The plate was washed, then incubated with 50 ul per well of a mouse anti-human IFNG biotinylated antibody (Cat. No. M701B) in quadruplicate at a concentration of 0.05 ug/ml for 2 hours at room temperature. The plate was washed and incubated with 100 ul per well of Streptavidin-HRP (Cat. No. N504) in all test wells at 1:40,000 for 30 minutes at room temperature. Detection was performed using 1-Step Ultra TMB Substrate (Cat. No. 34028) for 30 minutes at room temperature in the dark. The plate was then stopped with 0.16M sulfuric acid. Absorbances were read on a spectrophotometer at 450-550 nm.
Annotated product references
Cat. No. PA1-036 was used in immunohistochemistry to study the protective effect of cannabidiol against renal ischemia/reperfusion injury in rats. Fouad AA, Al-Mulhim AS, Jresat I (2012) Cannabidiol treatment ameliorates ischemia/reperfusion renal injury in rats. Life Sci 91:284–292.
Cat. No. PA1-036 was used in western blot to study the role of the expression profile of iNOS and arginase in rat macrophages in their resistance to Toxoplasma gondii infection. Li Z, Zhao ZJ, Zhu XQ et al. (2012) Differences in iNOS and arginase expression and activity in the macrophages of rats are responsible for the resistance against T. gondii infection. PLoS One 7:e35834.
Cat. No. MA1-516 was used in immunohistochemistry and western blot to study the role of HIF-1 alpha in the upregulation of HIF-1 beta in hypoxic human melanoma cells. Mandl M, Kapeller B, Lieber R et al. (2013) Hypoxia-inducible factor 1β (HIF-1β) is upregulated in a HIF-1α-dependent manner in 518A2 human melanoma cells under hypoxic conditions. Biochem Biophys Res Commun 434:166–172.
Cat. No. MA1-516 was used in western blot to study the role of HIF-1 alpha and VEGF in the neuroprotective effects of soy following acute stroke. Ma Y, Lovekamp-Swan T, Bekele W et al. (2013) Hypoxia-inducible factor and vascular endothelial growth factor are targets of dietary soy during acute stroke in female rats. Endocrinology 154:1589–1597.
Cat. No. M701B was used in ELISA to study the role of human melanoma cell Wnt/beta-catenin signaling in immune suppression and resistance. Yaguchi T, Goto Y, Kido K et al. (2012) Immune suppression and resistance mediated by constitutive activation of Wnt/β-catenin signaling in human melanoma cells. JImmunol 185:2110−2117.
Cat. No. M701B was used in ELISA to develop and validate novel bispecific antibodies that activate and direct T cells against tumor cells. Cui H, Thomas JD, Burke TR Jr et al. (2012) Chemically programmed bispecific antibodies that recruit and activate T cells. J Biol Chem 287:28206−28214.
For Research Use Only. Not for use in diagnostic procedures.