Western blot analysis was performed on nuclear enriched extracts (30 µg lysate) of HEL 92.1.7 (Lane 1), HL-60 (Lane 2) and K-562 (Lane 3). The blot was probed with Anti-TRIM28 Mouse Monoclonal Antibody (Product # 730029, 2 µg/ml) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjµgate (Product # A28177, 0.4 µg/ml, 1:2500 dilution). A ~88 kDa band corresponding to TRIM28 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody using iBind™ Flex Western Starter Kit (Product# SLF2000S). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG|
|Immunogen||619-811 of 835 of TRIM28 Protein.|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||2 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.
|Immunoprecipitation (IP)||See 1 publications below|
The protein encoded by this gene mediates transcriptional control by interaction with the Kruppel-associated box repression domain found in many transcription factors. The protein localizes to the nucleus and is thought to associate with specific chromatin regions. The protein is a member of the tripartite motif family. This tripartite motif includes three zinc-binding domains, a RING, a B-box type 1 and a B-box type 2, and a coiled-coil region.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Assessment of a method to characterize antibody selectivity and specificity for use in immunoprecipitation.
730029 was used in immunoprecipitation to describe a mass spectrometry-based method for scoring immunoprecipitation antibody quality
|Marcon E,Jain H,Bhattacharya A,Guo H,Phanse S,Pu S,Byram G,Collins BC,Dowdell E,Fenner M,Guo X,Hutchinson A,Kennedy JJ,Krastins B,Larsen B,Lin ZY,Lopez MF,Loppnau P,Miersch S,Nguyen T,Olsen JB,Paduch M,Ravichandran M,Seitova A,Vadali G,Vogelsang MS,Whiteaker JR,Zhong G,Zhong N,Zhao L,Aebersold R,Arrowsmith CH,Emili A,Frappier L,Gingras AC,Gstaiger M,Paulovich AG,Koide S,Kossiakoff AA,Sidhu SS,Wodak SJ,Gräslund S,Greenblatt JF,Edwards AM||Nature methods (12:725)||2015|