Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y320] to Tuberin
- Suitable for: WB, IHC-P, Flow Cyt
- Knockout validated
- Reacts with: Mouse, Rat, Human
Product nameAnti-Tuberin antibody [Y320]
See all Tuberin primary antibodies
DescriptionRabbit monoclonal [Y320] to Tuberin
Specificityab32554 recognises Tuberous sclerosis complexe-2 (TCS2 also known as Tuberin. It is predicted to detect isoform 2-4 based on sequence homology.
Tested applicationsSuitable for: WB, IHC-P, Flow Cytmore details
Species reactivityReacts with: Mouse, Rat, Human
Synthetic peptide within Human Tuberin aa 1750-1850 (C terminal). The exact sequence is proprietary.
- WB: HAP1 and HeLa whole cell lysates. Rat and mouse brain tissue lysates; Flow Cyt: Jurkat cells; IHC: Rat and mouse liver tissue. Human lung carcinoma tissue.
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol, 0.05% BSA
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab32554 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000 - 1/10000. Detects a band of approximately 220 kDa (predicted molecular weight: 200 kDa).|
|IHC-P||1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
For unpurified use at 1/100 dilution
FunctionIn complex with TSC1, inhibits the nutrient-mediated or growth factor-stimulated phosphorylation of S6K1 and EIF4EBP1 by negatively regulating mTORC1 signaling. Acts as a GTPase-activating protein (GAP) for the small GTPase RHEB, a direct activator of the protein kinase activity of mTORC1. Implicated as a tumor suppressor. Involved in microtubule-mediated protein transport, but this seems to be due to unregulated mTOR signaling. Stimulates weakly the intrinsic GTPase activity of the Ras-related proteins RAP1A and RAB5 in vitro. Mutations in TSC2 lead to constitutive activation of RAP1A in tumors.
Tissue specificityLiver, brain, heart, lymphocytes, fibroblasts, biliary epithelium, pancreas, skeletal muscle, kidney, lung and placenta.
Involvement in diseaseDefects in TSC2 are the cause of tuberous sclerosis type 2 (TSC2) [MIM:613254]. TSC2 is an autosomal dominant multi-system disorder that affects especially the brain, kidneys, heart, and skin. It is characterized by hamartomas (benign overgrowths predominantly of a cell or tissue type that occurs normally in the organ) and hamartias (developmental abnormalities of tissue combination). Clinical symptoms can range from benign hypopigmented macules of the skin to profound mental retardation with intractable seizures to premature death from a variety of disease-associated causes.
Defects in TSC2 are a cause of lymphangioleiomyomatosis (LAM) [MIM:606690]. LAM is a progressive and often fatal lung disease characterized by a diffuse proliferation of abnormal smooth muscle cells in the lungs. It affects almost exclusively young women and can occur as an isolated disorder or in association with tuberous sclerosis complex.
Sequence similaritiesContains 1 Rap-GAP domain.
modificationsPhosphorylation at Ser-1387, Ser-1418 or Ser-1420 does not affect interaction with TSC1.
Phosphorylation at Ser-939 and Thr-1462 by PKB/AKT1 is induced by growth factor stimulation.
Cellular localizationCytoplasm. Membrane. At steady state found in association with membranes.
- Information by UniProt
- FLJ43106 antibody
- LAM antibody
- OTTHUMP00000158940 antibody
All lanes : Anti-Tuberin antibody [Y320] (ab32554) at 1/5000 dilution (purified)
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate boiled
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate unboiled
Lane 3 : Mouse brain lysate boiled
Lane 4 : Mouse brain lysate unboiled
Lane 5 : Rat brain lysate boiled
Lane 6 : Rat brain lysate unboiled
Lysates/proteins at 20 µg per lane.
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 200 kDa
Observed band size: 200 kDa
The unboiled lysate are recommended to be used in SDS-PAGE, in order to prevent membrane protein aggregation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat liver tissue sections labeling Tuberin with purified ab32554 at 1/100 dilution (2.97 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labeling Tuberin with purified ab32554 at 1/100 dilution (2.97 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue sections labeling Tuberin with purified ab32554 at 1/100 dilution (2.97 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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 - unpurified ab32554 observed at 180 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab32554 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. Ab32554 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 ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling Tuberin with purified ab32554 at 1/30 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab32554 has been referenced in 16 publications.
- Zhang X et al. Tanshinone IIA protects against heart failure post-myocardial infarction via AMPKs/mTOR-dependent autophagy pathway. Biomed Pharmacother 112:108599 (2019). PubMed: 30798134
- Wang H et al. Feedback Activation of SGK3 and AKT Contributes to Rapamycin Resistance by Reactivating mTORC1/4EBP1 Axis via TSC2 in Breast Cancer. Int J Biol Sci 15:929-941 (2019). PubMed: 31182914
- Hadley G et al. Investigation of the novel mTOR inhibitor AZD2014 in neuronal ischemia. Neurosci Lett 706:223-230 (2019). PubMed: 31100427
- Ruvolo PP et al. Role of MSC-derived galectin 3 in the AML microenvironment. Biochim Biophys Acta 1865:959-969 (2018). PubMed: 29655803
- Struchtrup A et al. The ciliary protein RPGRIP1L governs autophagy independently of its proteasome-regulating function at the ciliary base in mouse embryonic fibroblasts. Autophagy 14:567-583 (2018). PubMed: 29372668