Immunofluorescence analysis of Tuberin / TSC2 was performed using 70% confluent log phase MCF-7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with Tuberin / TSC2 (7249) Mouse Monoclonal Antibody (730014) 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) 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||Human|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG|
|Immunogen||1525-1742 of 1807 of TSC2 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|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunoprecipitation (IP)||2 ug|
* 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|
Mutations in TSC2 lead to tuberous sclerosis complex. The protein is believed to be a tumor suppressor and is able to specifically stimulate the intrinsic GTPase activity of the Ras-related protein RAP1A and RAB5. The protein associates with hamartin in a cytosolic complex, possibly acting as a chaperone for hamartin. TSC2 may have a function in vesicular transport, but may also play a role in the regulation of cell growth arrest and in the regulation of transcription mediated by steroid receptors. Interaction between TSC1 and TSC2 may facilitate vesicular docking.
IP-MS enrichment of TSC2 (LFQ intensity): TSC2 was enriched 81-fold from HCT116 lysate compared to background proteins, using the optimized IP-MS workflow with Pierce MS-Compatible Magnetic IP Kit protein A/G (Part No. 90409) and TSC2 antibody (Part No. 730014). See more information on IP-MS verification of antibody selectivity. IP-MS validation info.
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.
730014 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|