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Ribosomal RNA (RRNA 5.8s) Antikörper

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Klonalität (Klon)
Monoklonal ()
Immunocytochemistry (ICC), Immunofluorescence (IF), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunoprecipitation (IP), Immunohistochemistry (IHC)
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Immunogen The whole 5.8S ribosomal RNA.
Klon Y10b
Isotyp IgG3 kappa
Spezifität This is specific for ribosomal RNA. There is no cross-reactivity with other RNAs.
Reinigung Protein A purified
Hintergrund The inhibition of protein synthesis by specific anti 5.8S rRNA oligonucleotides havesuggested that this RNA plays an important role in eukaryotic ribosome function,specifically ribosomal translocation. Mutations in the 5.8S rRNA can inhibit cell growthand compromise protein synthesis in vitro. This antibody is useful for recognizing RNA epitopes on both the large and smallsubunit. Alternate Names: anti-ribosomal RNA 5.8s antibody.
Applikationshinweise This rRNA Antibody (Y10b) is useful for Immunocytochemistry/Immunofluorescence and Immunoprecipitation. In ICC/IF, staining was observed in Ribosomes in the nucleoli and cytoplasm of Ntera2 cells.
Recommended dilutions: Immunocytochemistry/Immunofluorescence 1:2500, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunoprecipitation 10 µL/lysate
Protokoll Immunocytochemistry Protocol Specific for rRNA Antibody (Y10b) Immunocytochemistry ProtocolCulture cells to appropriate density in 35 mm culture dishes or 6-well plates.
1. Remove culture medium and add 10 % formalin to the dish. Fix at room temperature for 30 minutes.
. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
. To block nonspecific antibody binding incubate in 10 % normal goat serum from 1 hour to overnight at room temperature.
. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.
Beschränkungen Nur für Forschungszwecke einsetzbar
Format Liquid
Konzentration 1 mg/mL
Buffer 1 M Tris, 1.1 M NaCl, 100 mM Glycine, Sodium Azide
Konservierungsmittel Sodium azide
Vorsichtsmaßnahmen WARNING: Reagents contain sodium azide. Sodium azide is very toxic if ingested or inhaled. Avoid contact with skin, eyes, or clothing. Wear eye or face protection when handling. If skin or eye contact occurs, wash with copious amounts of water. If ingested or inhaled, contact a physician immediately. Sodium azide yields toxic hydrazoic acid under acidic conditions. Dilute azide-containing compounds in running water before discarding to avoid accumulation of potentially explosive deposits in lead or copper plumbing.
Handhabung Do not freeze.
Lagerung 4 °C
Produkt verwendet in: Jablonka, Wiese, Sendtner: "Axonal defects in mouse models of motoneuron disease." in: Journal of neurobiology, Vol. 58, Issue 2, pp. 272-86, 2004

Anderson, Merhege, Morin, Bolognani, Perrone-Bizzozero: "Increased expression and localization of the RNA-binding protein HuD and GAP-43 mRNA to cytoplasmic granules in DRG neurons during nerve regeneration." in: Experimental neurology, Vol. 183, Issue 1, pp. 100-8, 2003

Stone, Rubel: "Temporal, spatial, and morphologic features of hair cell regeneration in the avian basilar papilla." in: The Journal of comparative neurology, Vol. 417, Issue 1, pp. 1-16, 2000

Gallouzi, Brennan, Stenberg, Swanson, Eversole, Maizels, Steitz: "HuR binding to cytoplasmic mRNA is perturbed by heat shock." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 97, Issue 7, pp. 3073-8, 2000

Allgemeine Veröffentlichungen Bleher, Martin: "Ribosomes in the squid giant axon." in: Neuroscience, Vol. 107, Issue 3, pp. 527-34, 2001

Fan, Steitz: "Overexpression of HuR, a nuclear-cytoplasmic shuttling protein, increases the in vivo stability of ARE-containing mRNAs." in: The EMBO journal, Vol. 17, Issue 12, pp. 3448-60, 1998