Antibody data
- Antibody Data
- Antigen structure
- References [1]
- Comments [0]
- Validations
- Immunocytochemistry [1]
- Flow cytometry [1]
- Other assay [1]
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Validation data
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- Product number
- 730014 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Anti-TSC2 Monoclonal Antibody (7249)
- Antibody type
- Monoclonal
- Antigen
- Other
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 7249
- Vial size
- 100 µg
- Concentration
- 0.5 mg/ml
- Storage
- Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C
Submitted references Assessment of a method to characterize antibody selectivity and specificity for use in immunoprecipitation.
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 2015 Aug;12(8):725-31
Nature methods 2015 Aug;12(8):725-31
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- 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 (Product # 730014) 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 (Product # 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 (Product # 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.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometry analysis of Tuberin / TSC2 was done on MCF7 cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with Tuberin Mouse Monoclonal Antibody (730014, red histogram) or with mouse isotype control (pink histogram) at 3-5 ug/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Rabbit Anti-Mouse Secondary Antibody (A11059) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- 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 (Product # 90409) and TSC2 antibody (Product # 730014). See more information on IP-MS verification of antibody selectivity.