Immunofluorescence analysis of TIF1 beta 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 TRIM28 (20C1) Mouse Monoclonal Antibody (MA12023) at 2ug/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 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Mouse|
|Host / Isotype||Mouse / IgG|
|Immunogen||Recombinant protein corresponding to residues 60-383 of the human TIF1 beta protein.|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||Assay Dependent|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Gel Shift (GS)||Assay Dependent|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||1 - 3 µg/ml|
|Immunocytochemistry (ICC)||See 1 publications below|
MA1-2023 detects human and mouse TIF1 beta.
MA1-2023 has been used successfully in Western blot, and immunofluorescence procedures. By Western blot this antibody detects ~100 kDa protein representing TIF1 beta in gamma irradiated HeLa cell lysate. In immunofluorescence procedures, MA1-2023 recognizes TIF1 beta in gamma irradiated HeLa cells.
The MA1-2023 immunogen is recombinant protein corresponding to residues 60-383 of the human TIF1 beta protein.
TIF1 beta is a member of the tripartite motif (TRIM) family. It is composed of a N-terminal RBCC (RING finger, B boxes, coiled coil) motif, and a C-terminal bromodomain which is preceded by a PHD finger. TIF1 beta has been reported to function as a co-repressor for Kruppel-associated box (KRAB)-domain-containing zinc finger proteins. As such, it plays a crucial role in early embryogenesis. It appears that the co-repressor function of TIF1 beta is related to its organization and/or maintenance of higher-order, heterochromatin-like structures.
TRIM28 is required by the mouse KRAB domain protein ZFP568 to control convergent extension and morphogenesis of extra-embryonic tissues.
MA1-2023 was used in immunocytochemistry to investigate the effect of TRIM28 on extra-embryonic tissue development
|Shibata M,Blauvelt KE,Liem KF,García-García MJ||Development (Cambridge, England) (138:5333)||2011|