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Immunofluorescence analysis of Ribosomal RNA was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Ribosomal RNA 5.8S (Y10B) Mouse Monoclonal Antibody (MA113017) at 2µg/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Many|
|Host / Isotype||Mouse / IgG3, kappa|
|Immunogen||Whole ribosomal rRNA 5.8S|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunoprecipitation (IP)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
MA1-13017 detects ribosomal RNA 5.8S in many species.
MA1-13017 has been successfully used in Immunocytochemistry, immunoprecipitation, immunofluorescence applications.
The MA1-13017 immunogen is from whole ribosomal rRNA 52.8S
The inhibition of protein synthesis by specific anti 5.8S rRNA oligonucleotides have suggested that this RNA plays an important role in eukaryotic ribosome function, specifically ribosomal translocation. Mutations in the 5.8S rRNA can inhibit cell growth and compromise protein synthesis in vitro. This antibody is useful for recognizing RNA epitopes on both the large and small subunit.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.