Anti-Tuberin antibody [EP1107Y] - BSA and Azide free (ab247339)

Knockout Tested Rabbit Recombinant Monoclonal Tuberin antibody. Carrier free. Suitable for IHC-P, IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human samples.

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Images

Western blot - Anti-Tuberin antibody [EP1107Y] - BSA and Azide free (AB247339), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

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Product promiseTestedExpectedPredictedNot recommended

	IHC-P	IP	WB	ICC/IF	Flow Cyt (Intra)
Human	Tested	Tested	Tested	Tested	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Associated Products

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Target data

See full target information TSC2

Function

Catalytic component of the TSC-TBC complex, a multiprotein complex that acts as a negative regulator of the canonical mTORC1 complex, an evolutionarily conserved central nutrient sensor that stimulates anabolic reactions and macromolecule biosynthesis to promote cellular biomass generation and growth (PubMed:12172553, PubMed:12271141, PubMed:12842888, PubMed:12906785, PubMed:15340059, PubMed:22819219, PubMed:24529379, PubMed:28215400, PubMed:33436626, PubMed:35772404). Within the TSC-TBC complex, TSC2 acts as a GTPase-activating protein (GAP) for the small GTPase RHEB, a direct activator of the protein kinase activity of mTORC1 (PubMed:12172553, PubMed:12820960, PubMed:12842888, PubMed:12906785, PubMed:15340059, PubMed:22819219, PubMed:24529379, PubMed:33436626). In absence of nutrients, the TSC-TBC complex inhibits mTORC1, thereby preventing phosphorylation of ribosomal protein S6 kinase (RPS6KB1 and RPS6KB2) and EIF4EBP1 (4E-BP1) by the mTORC1 signaling (PubMed:12172553, PubMed:12271141, PubMed:12842888, PubMed:12906785, PubMed:22819219, PubMed:24529379, PubMed:28215400, PubMed:35772404). The TSC-TBC complex is inactivated in response to nutrients, relieving inhibition of mTORC1 (PubMed:12172553, PubMed:24529379). Involved in microtubule-mediated protein transport via its ability to regulate mTORC1 signaling (By similarity). Also stimulates the intrinsic GTPase activity of the Ras-related proteins RAP1A and RAB5 (By similarity).

Alternative names

TSC4, TSC2, Tuberin, Tuberous sclerosis 2 protein

Recommended products

Knockout Tested Rabbit Recombinant Monoclonal Tuberin antibody. Carrier free. Suitable for IHC-P, IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human samples.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
Clone number: EP1107Y
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: +4°C
Storage information: Do Not Freeze

Notes

ab247339 is the carrier-free version of Anti-Tuberin antibody [EP1107Y] ab52936.

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Tuberin also known as TSC2 is a tumor suppressor protein with a role in cell growth regulation. It has a molecular mass of approximately 200 kDa. This protein forms a critical component of the TSC1-TSC2 complex and is expressed widely in various tissues including the brain kidney heart and skin. Tuberin possesses GTPase-activating protein (GAP) activity targeting specifically the small GTPase protein Rheb which leads to inhibition of the mammalian target of rapamycin complex 1 (mTORC1).

Biological function summary

Tuberin helps control cell size and proliferation. It does this by modulating the mTOR signaling pathway. This modulation is essential for maintaining cellular homeostasis and energy balance particularly under conditions of nutrient scarcity. Tuberin as part of the TSC1-TSC2 complex also influences cell cycle progression and protein synthesis. Its functions are vital in preventing abnormal cell growth and division which is particularly important in organs where rapid cell turnover occurs.

Pathways

Tuberin plays a central role in the mTOR signaling pathway which affects cell growth autophagy and metabolism. It directly interacts with hamartin (TSC1) to form the TSC1-TSC2 complex which regulates Rheb activity. By inhibiting Rheb Tuberin helps control the mTORC1 pathway linking it to signals such as growth factors stress energy status and amino acid availability. This coordination is important for the proper balance between cell growth and catabolism.

Associated diseases and disorders

Tuberin is mainly associated with Tuberous Sclerosis Complex (TSC) and certain forms of epilepsy. Mutations or loss of function in TSC2 can lead to benign tumor formations in multiple organs including the brain kidneys and skin. It is also implicated in the development of lymphangioleiomyomatosis (LAM) a rare lung disease. Tuberin's interaction with mTORC1 highlights its connection with these conditions as hyperactivation of the mTOR pathway contributes significantly to the pathogenesis.

Product promise

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9 product images

Western blot - Anti-Tuberin antibody [EP1107Y] - BSA and Azide free (ab247339)
This data was developed using Anti-Tuberin antibody [EP1107Y] ab52936, the same antibody clone in a different buffer formulation.
Lane 1: Wild-type HAP1 whole cell lysate (20 μg)
Lane 2: Tuberin knockout HAP1 whole cell lysate (20 μg)
Lane 3: HeLa whole cell lysate (20 μg)
Lanes 1 - 4: Merged signal (red and green). Green - Anti-Tuberin antibody [EP1107Y] ab52936 observed at 180 kDa. Red - loading control, ab18058, observed at 130 kDa.
Anti-Tuberin antibody [EP1107Y] ab52936 was shown to specifically react with tuberin in wild-type HAP1 cells. No band was observed when tuberin knockout samples were used. Wild-type and tuberin knockout samples were subjected to SDS-PAGE. Anti-Tuberin antibody [EP1107Y] ab52936 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Tuberin antibody [EP1107Y] (Anti-Tuberin antibody [EP1107Y] ab52936)
Predicted band size: 201 kDa
Immunocytochemistry/ Immunofluorescence - Anti-Tuberin antibody [EP1107Y] - BSA and Azide free (ab247339)
This data was developed using Anti-Tuberin antibody [EP1107Y] ab52936, the same antibody clone in a different buffer formulation.Anti-Tuberin antibody [EP1107Y] ab52936 stained HeLa cells. The cells were 4% formaldehyde fixed for 10 minutes at room temperature and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (Anti-Tuberin antibody [EP1107Y] ab52936 at 1/100 dilution) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
Western blot - Anti-Tuberin antibody [EP1107Y] - BSA and Azide free (ab247339)
This data was developed using Anti-Tuberin antibody [EP1107Y] ab52936, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-Tuberin antibody [EP1107Y] (Anti-Tuberin antibody [EP1107Y] ab52936) at 1/20000 dilution
All lanes: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate prepared in 1%SDS Hot lysis method at 15 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 201 kDa
Observed band size: 200 kDa
Immunoprecipitation - Anti-Tuberin antibody [EP1107Y] - BSA and Azide free (ab247339)
This data was developed using Anti-Tuberin antibody [EP1107Y] ab52936, the same antibody clone in a different buffer formulation.Tuberin was immunoprecipitated from 0.35 mg HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg with Anti-Tuberin antibody [EP1107Y] ab52936 at 1/50 dilution (2µg). VeriBlot for IP Detection Reagent (HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg
Lane 2: abab52936 IP in HeLa whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Tuberin antibody [EP1107Y] ab52936 in HeLa whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-Tuberin antibody [EP1107Y] (Anti-Tuberin antibody [EP1107Y] ab52936)
Predicted band size: 201 kDa
Observed band size: 200 kDa
Flow Cytometry (Intracellular) - Anti-Tuberin antibody [EP1107Y] - BSA and Azide free (ab247339)
This data was developed using Anti-Tuberin antibody [EP1107Y] ab52936, the same antibody clone in a different buffer formulation.Overlay histogram showing HeLa cells stained with Anti-Tuberin antibody [EP1107Y] ab52936 (red line). The cells were fixed with 80% methanol (5 min)/ and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (Anti-Tuberin antibody [EP1107Y] ab52936, 1/10000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1?g/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HeLa cells fixed with 4% formaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Western blot - Anti-Tuberin antibody [EP1107Y] - BSA and Azide free (ab247339)
This data was developed using Anti-Tuberin antibody [EP1107Y] ab52936, the same antibody clone in a different buffer formulation.
The cell lysates was prepared in 1%SDS Hot lysis method.
All lanes: Western blot - Anti-Tuberin antibody [EP1107Y] (Anti-Tuberin antibody [EP1107Y] ab52936) at 1/100000 dilution
All lanes: SH-SY5Y cell lysate at 10 µg
Secondary
All lanes: HRP-labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 201 kDa
Observed band size: ~200 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tuberin antibody [EP1107Y] - BSA and Azide free (ab247339)
This data was developed using Anti-Tuberin antibody [EP1107Y] ab52936, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded human adenocarcinoma of uterus using Anti-Tuberin antibody [EP1107Y] ab52936 at a dilution of 1/100-1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cytometry (Intracellular) - Anti-Tuberin antibody [EP1107Y] - BSA and Azide free (ab247339)
This data was developed using Anti-Tuberin antibody [EP1107Y] ab52936, the same antibody clone in a different buffer formulation.
Overlay histogram showing SH-SY5Y (Human neuroblastoma epithelial cell) cells stained with Anti-Tuberin antibody [EP1107Y] ab52936 (red line). The cells were fixed with 4% Paraformaldehyde and then permeabilized with 90% Methanol. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H&L) Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 at 1/2000 dilution. Isotype control antibody (black line) was rabbit IgG (monoclonal). Unlabelled sample (blue line) was Cell without incubation with primary antibody and secondary antibody.
Western blot - Anti-Tuberin antibody [EP1107Y] - BSA and Azide free (ab247339)
Western blot: Anti-TSC2 antibody [EP1107Y] (Anti-Tuberin antibody [EP1107Y] ab52936) staining at 1/50000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (Anti-Calnexin antibody [CANX/1543] ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-Tuberin antibody [EP1107Y] ab52936 was shown to bind specifically to TSC2. A band was observed at 200 kDa in wild-type A549 cell lysates with no signal observed at this size in TSC2 knockout cell line. To generate this image, wild-type and TSC2 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-Tuberin antibody [EP1107Y] (Anti-Tuberin antibody [EP1107Y] ab52936) at 1/50000 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: TSC2 knockout A549 cell lysate at 20 µg
Lane 3: Wild-type HAP1 cell lysate at 20 µg
Lane 4: TSC2 knockout HAP1 cell lysate at 20 µg
Secondary
All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.