Validated using a knockout cell lineRecombinantRabMAb

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

Submit a review Submit a question
Western blot - Anti-Tuberin antibody [EP1107Y] - BSA and Azide free (ab247339)
  • Immunocytochemistry - Anti-Tuberin antibody [EP1107Y] - BSA and Azide free (ab247339)
  • Western blot - Anti-Tuberin antibody [EP1107Y] - BSA and Azide free (ab247339)
  • Flow Cytometry - Anti-Tuberin antibody [EP1107Y] - BSA and Azide free (ab247339)
  • Flow Cytometry - Anti-Tuberin antibody [EP1107Y] - BSA and Azide free (ab247339)
  • Western blot - Anti-Tuberin antibody [EP1107Y] - BSA and Azide free (ab247339)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tuberin antibody [EP1107Y] - BSA and Azide free (ab247339)
  • Anti-Tuberin antibody [EP1107Y] - BSA and Azide free (ab247339)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1107Y] to Tuberin - BSA and Azide free
  • Suitable for: IHC-P, WB, ICC/IF, Flow Cyt, IP
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-Tuberin antibody [EP1107Y] - BSA and Azide free
    See all Tuberin primary antibodies

  • Description

    Rabbit monoclonal [EP1107Y] to Tuberin - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, WB, ICC/IF, Flow Cyt, IPmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide within Human Tuberin aa 1750-1850. The exact sequence is proprietary.

  • General notes

    ab247339 is the carrier-free version of ab52936 This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    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.

    Ab247339 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

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

    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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab247339 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
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB Use at an assay dependent concentration. Detects a band of approximately 201 kDa (predicted molecular weight: 201 kDa).
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.
IP Use at an assay dependent concentration.

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

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

      This data was developed using 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 - ab52936 observed at 180 kDa. Red - loading control, ab18058, observed at 130 kDa.

      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. 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 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.

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

      This data was developed using ab52936, the same antibody clone in a different buffer formulation.

      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 (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 (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)
      Western blot - Anti-Tuberin antibody [EP1107Y] - BSA and Azide free (ab247339)
      Anti-Tuberin antibody [EP1107Y] (ab52936) at 1/20000 dilution + HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate prepared in 1%SDS Hot lysis method at 15 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 201 kDa
      Observed band size: 200 kDa
      why is the actual band size different from the predicted?



      This data was developed using ab52936, the same antibody clone in a different buffer formulation.

      Blocking and dilution buffer: 5% NFDM/TBST.

    • Flow Cytometry - Anti-Tuberin antibody [EP1107Y] - BSA and Azide free (ab247339)
      Flow Cytometry - Anti-Tuberin antibody [EP1107Y] - BSA and Azide free (ab247339)
      This data was developed using ab52936, the same antibody clone in a different buffer formulation.Overlay histogram showing SH-SY5Y (Human neuroblastoma epithelial cell) cells stained with 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) 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.
    • Flow Cytometry - Anti-Tuberin antibody [EP1107Y] - BSA and Azide free (ab247339)
      Flow Cytometry - Anti-Tuberin antibody [EP1107Y] - BSA and Azide free (ab247339)
      This data was developed using ab52936, the same antibody clone in a different buffer formulation.Overlay histogram showing HeLa cells stained with 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 (ab52936, 1/10000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (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)
      Western blot - Anti-Tuberin antibody [EP1107Y] - BSA and Azide free (ab247339)
      Anti-Tuberin antibody [EP1107Y] (ab52936) at 1/100000 dilution + SH-SY5Y cell lysate at 10 µg

      Secondary
      HRP-labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 201 kDa
      Observed band size: ~200 kDa why is the actual band size different from the predicted?



      This data was developed using ab52936, the same antibody clone in a different buffer formulation.

      The cell lysates was prepared in 1%SDS Hot lysis method.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tuberin antibody [EP1107Y] - BSA and Azide free (ab247339)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tuberin antibody [EP1107Y] - BSA and Azide free (ab247339)
      This data was developed using ab52936, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded human adenocarcinoma of uterus using 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.
    • Anti-Tuberin antibody [EP1107Y] - BSA and Azide free (ab247339)
      Anti-Tuberin antibody [EP1107Y] - BSA and Azide free (ab247339)

    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 (0)

    Publishing research using ab247339? Please let us know so that we can cite the reference in this datasheet.

    ab247339 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    There are currently no Customer reviews or Questions for ab247339.
    Please use the links above to contact us or submit feedback about this product.

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com