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AB248510

Anti-Tuberin (phospho S664) antibody [EPR8202] - BSA and Azide free

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Rabbit Recombinant Monoclonal Tuberin phospho S664 antibody. Carrier free. Suitable for WB and reacts with Human, Mouse, Rat samples.

View Alternative Names

TSC4, TSC2, Tuberin, Tuberous sclerosis 2 protein

5 Images
Western blot - Anti-Tuberin (phospho S664) antibody [EPR8202] - BSA and Azide free (AB248510)
  • WB

Unknown

Western blot - Anti-Tuberin (phospho S664) antibody [EPR8202] - BSA and Azide free (AB248510)

This data was developed using ab133465, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-Tuberin (phospho S664) antibody [EPR8202] (<a href='/en-us/products/primary-antibodies/tuberin-phospho-s664-antibody-epr8202-ab133465'>ab133465</a>) at 1/1000 dilution

Lane 1:

SH SY5Y cell lysates at 10 µg

Lane 2:

SH SY5Y cell lysates treated with Okadaic acid and Calyculin A at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 201 kDa

false

Western blot - Anti-Tuberin (phospho S664) antibody [EPR8202] - BSA and Azide free (AB248510)
  • WB

Lab

Western blot - Anti-Tuberin (phospho S664) antibody [EPR8202] - BSA and Azide free (AB248510)

Blocking/Diluting buffer and concentration 5% NFDM/TBST This data was developed using ab133465, the same antibody clone in a different buffer formulation.

Lanes 1 - 3:

Western blot - Anti-Tuberin (phospho S664) antibody [EPR8202] (<a href='/en-us/products/primary-antibodies/tuberin-phospho-s664-antibody-epr8202-ab133465'>ab133465</a>) at 1/1000 dilution

Lanes 1 - 3:

Western blot - Anti-Tuberin (phospho S664) antibody [EPR8202] - BSA and Azide free (ab248510)

Lane 1:

Untreated HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate unboiled at 15 µg

Lane 2:

HeLa treated with 200nM Phorbol-12-myristate-13-acetate (PMA) for 4 hours whole cell lysate unboiled at 15 µg

Lane 3:

HeLa treated with 200nM Phorbol-12-myristate-13-acetate (PMA) for 4 hours whole cell lysate unboiled, then the membrane treated with Alkaline Phosphatase for 1 hour at 15 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 200 kDa

false

Exposure time: 80s

Western blot - Anti-Tuberin (phospho S664) antibody [EPR8202] - BSA and Azide free (AB248510)
  • WB

Lab

Western blot - Anti-Tuberin (phospho S664) antibody [EPR8202] - BSA and Azide free (AB248510)

Blocking/Diluting buffer and concentration 5% NFDM/TBST Exposure time : Lane 1, 2 : 180 seconds Lane 3, 4 : 40 seconds Lane 5, 6 : 20 seconds This data was developed using ab133465, the same antibody clone in a different buffer formulation.

Lanes 1 - 6:

Western blot - Anti-Tuberin (phospho S664) antibody [EPR8202] (<a href='/en-us/products/primary-antibodies/tuberin-phospho-s664-antibody-epr8202-ab133465'>ab133465</a>) at 1/1000 dilution

Lanes 1 - 6:

Western blot - Anti-Tuberin (phospho S664) antibody [EPR8202] - BSA and Azide free (ab248510)

Lane 1:

Human brain lysate unboiled at 15 µg

Lane 2:

Human brain lysate unboiled, then the membrane treated with Alkaline Phosphatase for 1 hour at 15 µg

Lane 3:

Rat brain lysate unboiled at 15 µg

Lane 4:

Rat brain lysate unboiled, then the membrane treated with Alkaline Phosphatase for 1 hour at 15 µg

Lane 5:

Mouse brain lysate unboiled at 15 µg

Lane 6:

Mouse brain lysate unboiled, then the membrane treated with Alkaline Phosphatase for 1 hour at 15 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 200 kDa

false

Western blot - Anti-Tuberin (phospho S664) antibody [EPR8202] - BSA and Azide free (AB248510)
  • WB

Lab

Western blot - Anti-Tuberin (phospho S664) antibody [EPR8202] - BSA and Azide free (AB248510)

Blocking/Diluting buffer and concentration 5% NFDM/TBST We recommend not to boil the samples after lysis to get desired WB results. The molecular weight observed is consistent with what has been described in the literature (PMID : 30774414; 10335945). This data was developed using ab133465, the same antibody clone in a different buffer formulation.

Lanes 1 - 4:

Western blot - Anti-Tuberin (phospho S664) antibody [EPR8202] (<a href='/en-us/products/primary-antibodies/tuberin-phospho-s664-antibody-epr8202-ab133465'>ab133465</a>) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Anti-Tuberin (phospho S664) antibody [EPR8202] - BSA and Azide free (ab248510)

Lane 1:

Mouse brain lysate boiled at 15 µg

Lane 2:

Mouse brain lysate unboiled at 15 µg

Lane 3:

Rat brain lysate boiled at 15 µg

Lane 4:

Rat brain lysate unboiled at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 200 kDa

false

Exposure time: 40s

OI-RD Scanning - Anti-Tuberin (phospho S664) antibody [EPR8202] - BSA and Azide free (AB248510)
  • OI-RD Scanning

Unknown

OI-RD Scanning - Anti-Tuberin (phospho S664) antibody [EPR8202] - BSA and Azide free (AB248510)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR8202

Isotype

IgG

Carrier free

Yes

Reacts with

Mouse, Rat, Human

Applications

WB

applications

Immunogen

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

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "FlowCyt" : {"fullname" : "Flow Cytometry", "shortname":"Flow Cyt"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "<p></p>", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "<p></p>", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "<p></p>", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "<p></p>" }, "Mouse": { "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "" }, "Rat": { "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "<p></p>", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "" } } }
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Product details

ab248510 is the carrier-free version of ab133465.

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

What are the advantages of a recombinant monoclonal antibody?
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.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. 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.

Compatibility
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.

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Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Constituents: PBS
Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze
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Supplementary information

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

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.

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.
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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
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Target data

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).
See full target information TSC2 phospho S664
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