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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Tuberin antibody [Y320] (ab32554)

Reviews (1)Q&A (2)References (18)
Western blot - Anti-Tuberin antibody [Y320] (ab32554)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tuberin antibody [Y320] (ab32554)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tuberin antibody [Y320] (ab32554)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tuberin antibody [Y320] (ab32554)
  • Western blot - Anti-Tuberin antibody [Y320] (ab32554)
  • Flow Cytometry (Intracellular) - Anti-Tuberin antibody [Y320] (ab32554)
  • Anti-Tuberin antibody [Y320] (ab32554)

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: Flow Cyt (Intra), WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

APC Carrier Free HRP PE

Overview

  • Product name

    Anti-Tuberin antibody [Y320]
    See all Tuberin primary antibodies

  • Description

    Rabbit monoclonal [Y320] to Tuberin

  • Host species

    Rabbit

  • Specificity

    ab32554 recognises Tuberous sclerosis complexe-2 (TCS2 also known as Tuberin. It is predicted to detect isoform 2-4 based on sequence homology.

  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IHC-Pmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human Tuberin aa 1750-1850 (C terminal). The exact sequence is proprietary.

  • Positive control

    • WB: HAP1 and HeLa whole cell lysates. Rat and mouse brain tissue lysates; Flow Cyt (intra): Jurkat cells; IHC: Rat and mouse liver tissue. Human lung carcinoma tissue.

  • General notes

    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
    For more information see here.

    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.

Properties

Applications

The Abpromise guarantee

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.

Application Abreviews Notes
Flow Cyt (Intra)
1/30.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.  For unpurified use at 1/100 dilution
WB (1)
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.
Notes
Flow Cyt (Intra)
1/30.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.  For unpurified use at 1/100 dilution
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.

Target

  • Function

    In 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 specificity

    Liver, brain, heart, lymphocytes, fibroblasts, biliary epithelium, pancreas, skeletal muscle, kidney, lung and placenta.

  • Involvement in disease

    Defects 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 similarities

    Contains 1 Rap-GAP domain.

  • Post-translational
    modifications

    Phosphorylation 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 localization

    Cytoplasm. Membrane. At steady state found in association with membranes.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • FLJ43106 antibody
    • LAM antibody
    • OTTHUMP00000158940 antibody
    • OTTHUMP00000198394 antibody
    • OTTHUMP00000198395 antibody
    • PPP1R160 antibody
    • Protein phosphatase 1, regulatory subunit 160 antibody
    • TSC complex subunit 2 antibody
    • tsc2 antibody
    • TSC2_HUMAN antibody
    • TSC4 antibody
    • TSC4 gene, formerly antibody
    • TSC4, formerly antibody
    • Tuberin antibody
    • Tuberous sclerosis 2 antibody
    • Tuberous sclerosis 2 protein antibody
    • Tuberous sclerosis 2 protein homolog antibody
    see all

Images

  • Western blot - Anti-Tuberin antibody [Y320] (ab32554)
    Western blot - Anti-Tuberin antibody [Y320] (ab32554)
    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.

    Secondary
    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) - Anti-Tuberin antibody [Y320] (ab32554)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tuberin antibody [Y320] (ab32554)

    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) - Anti-Tuberin antibody [Y320] (ab32554)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tuberin antibody [Y320] (ab32554)

    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) - Anti-Tuberin antibody [Y320] (ab32554)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tuberin antibody [Y320] (ab32554)

    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.

  • Western blot - Anti-Tuberin antibody [Y320] (ab32554)
    Western blot - Anti-Tuberin antibody [Y320] (ab32554)

    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 (Intracellular) - Anti-Tuberin antibody [Y320] (ab32554)
    Flow Cytometry (Intracellular) - Anti-Tuberin antibody [Y320] (ab32554)

    Intracellular Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling Tuberin with purified ab32554 at 1/30 dilution (10µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluorr® 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). 

     

     

  • Anti-Tuberin antibody [Y320] (ab32554)
    Anti-Tuberin antibody [Y320] (ab32554)

Protocols

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.

Click here to view the general protocols

References (18)

Publishing research using ab32554? Please let us know so that we can cite the reference in this datasheet.

ab32554 has been referenced in 18 publications.

  • Shiomi E  et al. Relationship between miR-155 expression and clear cell papillary renal cell carcinoma in the dialyzed kidney. IJU Case Rep 4:127-131 (2021). PubMed: 33977237
  • 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
View all Publications for this product

Customer reviews and Q&As

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1-3 of 3 Abreviews or Q&A

Western blot abreview for Anti-Tuberin antibody [Y320]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa)
Loading amount
20 µg
Specification
HeLa
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 45 minute(s) · Concentration: 5% · Temperature: 22°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Dr. jp lu

Verified customer

Submitted Nov 21 2008

Question

I have bought and measured samples with mTOR (Epitomics， 6585-1), Pi3k p85(Epitomics ,5405-1) and TSC2 antibody 1:1000(Epitomics，1613-1）, does that mean I have tested both non-phosphorylated and phosphorylated of mTOR, or Pi3k p85α or TSC2 respectly? Or these antibodys are just used for testing non-phosphorylated mTOR, Pi3k p85α or TSC2 respectly?

Read More
Answer

All three antibodies will detect the proteins with or without phosphorylation. The one exception may be the TSC2 antibody, which is raised against a C terminus peptide that contains a phosphorylation site at serine 1798. It is possible that if this amino acid is phosphorylated, the antibody will not bind to the protein, but we have not confirmed this. Here is a link to the UniProt entry for the human protein, which includes notes about the phosphorylation sites in the section titled Sequence annotations: http://www.uniprot.org/uniprot/P49815 .

Read More

Question

Buenas tardes,
Estaríamos interesados en los anticuerpos HAMARTINA y TUBERINA que sean de aplicación en inmunohistoquimia en parafina. Nos interesaría conocer carácter poli o monoclonal.
Por favor, necesitaríamos saber referencia , precio y plazo de entrega.
Esperamos con urgencia su respuesta.
Gracias,
Saludos

Read More
Answer

Muchas gracias por contactarnos.

Anticuerpos contra HAMARTINA, o TSC1 testados en IHC-P, tenemos cuatro, dos de conejo policlonal, uno monoclonal también de conejo, y uno policlonal de cabra. En función de las especies en las que funciona o podría funcionar, quizás os pueda interesar uno u otro. Adjunto la lista de los cuatro, y pinchando en cada uno de ellos se accede a su datasheet, donde encontrareis toda la información sobre el anticuerpo, y en la parte de arriba el precio y la disponibilidad.

https://www.abcam.com/index.html?pageconfig=searchresults&search=TSC1&pt=1&sk=app&sv=74&sn=IHC-P&l=2&fViewMore=1

Para TUBERINA, también caracterizados por IHC-P, las alternativas son mayores. Os animo a que echéis un vistazo a las fichas técnicas de cada uno de ellos, y me contactéis en caso de duda.

https://www.abcam.com/index.html?pageconfig=searchresults&search=Tuberin&pt=1&sk=app&sv=74&sn=IHC-P&l=2&fViewMore=1

Igualmente, el precio y la disponibilidad se pueden ver en la parte de arriba de la ficha técnica.

Si necesitáis un presupuesto, información adicional acerca de las aplicaciones testadas, o cualquier otra información, no dudéis en volverme a contactar (contestando a este mail o por teléfono en el 91 114 65 54).

Read More

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