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|
* 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.
|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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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|