Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online
Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Hamartin antibody [EP318Y] (ab40872)

Reviews (1)Q&A (3)References (3)
Western blot - Anti-Hamartin antibody [EP318Y] (ab40872)
  • Immunocytochemistry - Anti-Hamartin antibody [EP318Y] (ab40872)
  • Western blot - Anti-Hamartin antibody [EP318Y] (ab40872)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hamartin antibody [EP318Y] (ab40872)
  • Flow Cytometry (Intracellular) - Anti-Hamartin antibody [EP318Y] (ab40872)
  • Anti-Hamartin antibody [EP318Y] (ab40872)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP318Y] to Hamartin
  • Suitable for: WB, IHC-P, Flow Cyt (Intra), ICC/IF
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free

Overview

  • Product name

    Anti-Hamartin antibody [EP318Y]
    See all Hamartin primary antibodies

  • Description

    Rabbit monoclonal [EP318Y] to Hamartin

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt (Intra), ICC/IFmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • ICC: HeLa cells IHC: human liver tissue WB: HeLa cells; wild type HAP1 cell lysate, Hamartin knockout HAP1 cell lysate, HeLa cells, Human skeletal muscle tissue lysate Flow Cyt (intra): HeLa cells

  • 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 ab40872 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
WB
1/1000. Predicted molecular weight: 150 kDa.

For the unpurified version use 1/20000 dilution
IHC-P
1/200. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt (Intra)
1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.  For the unpurified version use 1/1000- 1/10000 dilution
ICC/IF
1/50.

For unpurified version use 1/500 dilution
Notes
WB
1/1000. Predicted molecular weight: 150 kDa.

For the unpurified version use 1/20000 dilution
IHC-P
1/200. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt (Intra)
1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.  For the unpurified version use 1/1000- 1/10000 dilution
ICC/IF
1/50.

For unpurified version use 1/500 dilution

Target

  • Function

    In complex with TSC2, inhibits the nutrient-mediated or growth factor-stimulated phosphorylation of S6K1 and EIF4EBP1 by negatively regulating mTORC1 signaling. Seems not to be required for TSC2 GAP activity towards RHEB. Implicated as a tumor suppressor. Involved in microtubule-mediated protein transport, but this seems to be due to unregulated mTOR signaling.

  • Tissue specificity

    Highly expressed in skeletal muscle, followed by heart, brain, placenta, pancreas, lung, liver and kidney. Also expressed in embryonic kidney cells.

  • Involvement in disease

    Defects in TSC1 are the cause of tuberous sclerosis type 1 (TSC1) [MIM:191100]. It is an autosomal dominant multi-system disorder that affects especially the brain, kidneys, heart, and skin. TS1C 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 TSC1 may be a cause of focal cortical dysplasia of Taylor balloon cell type (FCDBC) [MIM:607341]. FCDBC is a subtype of cortical displasias linked to chronic intractable epilepsy. Cortical dysplasias display a broad spectrum of structural changes, which appear to result from changes in proliferation, migration, differentiation, and apoptosis of neuronal precursors and neurons during cortical development.

  • Domain

    The C-terminal putative coiled-coil domain is necessary for interaction with TSC2.

  • Post-translational
    modifications

    Phosphorylation at Ser-505 does not affect interaction with TSC2. Phosphorylated upon DNA damage, probably by ATM or ATR.

  • Cellular localization

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

  • Database links

    • Alternative names

      • Hamartin antibody
      • kiaa0243 antibody
      • LAM antibody
      • TSC antibody
      • Tsc1 antibody
      • Tsc1 gene antibody
      • TSC1_HUMAN antibody
      • Tuberous sclerosis 1 antibody
      • Tuberous sclerosis 1 protein antibody
      • tumor suppressor antibody
      see all

    Images

    • Western blot - Anti-Hamartin antibody [EP318Y] (ab40872)
      Western blot - Anti-Hamartin antibody [EP318Y] (ab40872)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: Hamartin knockout HAP1 cell lysate (20 µg)
      Lane 3: HeLa cell lysate (20 µg)
      Lane 4: Human skeletal muscle tissue lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab40872 observed at 150 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab40872 was shown to recognize Hamartin when Hamartin knockout samples were used, along with additional cross-reactive bands. Wild-type and Hamartin knockout samples were subjected to SDS-PAGE. ab40872 and ab8245 (loading control to GAPDH) were diluted 1/5000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry - Anti-Hamartin antibody [EP318Y] (ab40872)
      Immunocytochemistry - Anti-Hamartin antibody [EP318Y] (ab40872)
      Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Hamartin with Purified ab40872 at 1:50 dilution (4.3 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488,ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
    • Western blot - Anti-Hamartin antibody [EP318Y] (ab40872)
      Western blot - Anti-Hamartin antibody [EP318Y] (ab40872)
      Anti-Hamartin antibody [EP318Y] (ab40872) at 1/1000 dilution + HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate

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

      Predicted band size: 150 kDa



      We are not sure about the nature of the extra band. 

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hamartin antibody [EP318Y] (ab40872)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hamartin antibody [EP318Y] (ab40872)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human liver tissue sections labeling Hamartin with purified ab40872 at 1/200 dilution (1.08 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). The immunostaining was performed on a Leica Biosystems BOND® RX instrument. 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.
    • Flow Cytometry (Intracellular) - Anti-Hamartin antibody [EP318Y] (ab40872)
      Flow Cytometry (Intracellular) - Anti-Hamartin antibody [EP318Y] (ab40872)

      Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Hamartin with Purified ab40872 at 1/20 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).

       

    • Anti-Hamartin antibody [EP318Y] (ab40872)
      Anti-Hamartin antibody [EP318Y] (ab40872)

    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 (3)

    Publishing research using ab40872? Please let us know so that we can cite the reference in this datasheet.

    ab40872 has been referenced in 3 publications.

    • Wang Y  et al. miR-130a upregulates mTOR pathway by targeting TSC1 and is transactivated by NF-?B in high-grade serous ovarian carcinoma. Cell Death Differ 24:2089-2100 (2017). PubMed: 28800130
    • Platt FM  et al. Spectrum of phosphatidylinositol 3-kinase pathway gene alterations in bladder cancer. Clin Cancer Res 15:6008-17 (2009). PubMed: 19789314
    • Pymar LS  et al. Bladder tumour-derived somatic TSC1 missense mutations cause loss of function via distinct mechanisms. Hum Mol Genet 17:2006-17 (2008). PubMed: 18397877

    Customer reviews and Q&As

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

    1-4 of 4 Abreviews or Q&A

    Flow Cytometry abreview for Anti-Hamartin antibody [EP318Y]

     Average
    Abreviews
    abreview image
    Application
    Flow Cytometry
    Sample
    Rabbit Cell (Human ESC)
    Permeabilization
    Yes - Several (see image)
    Gating Strategy
    Single cells (by scatter) and live cells (by scatter and Hoechst)
    Specification
    Human ESC
    Preparation
    Cell harvesting/tissue preparation method: Accutase and Resuspension
    Sample buffer: PBS
    Fixation
    Several (see image)
    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.

    Abcam user community

    Verified customer

    Submitted Aug 13 2019

    Question

    human, canc. kidney. high concentration 1/100.

    Read More
    Answer

    Vielen Dank für Ihre Anfrage und Ihre Geduld.

    Ich habe endlich Nachricht aus dem Labor erhalten, dass die Färbungen ein voller Erfolg waren. Die Bilder befinden sich im Anhang. Da die Färbung schön kräftig ausfällt, fürchte ich, dass der Antikörper beschädigt sein muss, den Sie erhalten haben. Wenn Sie möchten, sende ich ihnen gerne einen kostenlosen Ersatz zu.


    "I just received the news that this product performed well, in retest. We retested it in paraffin embedded tissues at dilutions of 50-100 and 250. Please see the attached images."


    Ich hoffe, dies hilft Ihnen weiter. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

    Benutzen Sie unsere Produkte? Schicken Sie uns einen Abreview. Verdienen Sie sich eine Belohnung!
    https://www.abcam.com/abreviews

    Read More

    Question

    besten Dank für Ihre Unterstützung.

    In der Anlage übersende ich das ausgefüllte Formular und sende auch Bilder dazu. Die Niere ist negativ, bei Lunge und Herz kann man eine schwache Reaktivität erkennen.

    Reagieren Erythrocyten mit dem AK? Man sieht zT auch intranukleäre globuläre Reaktivität.

    1) Abcam product code ab 40872

    2) Abcam order reference number or product batch number

    3) Description of the problem No specific positive staining using dilution of 1:50 and 1:100

    4) Sample preparation:
    Species
    Type of sample: Fresh frozen sections, perfusion fixed frozen sections, PFA/formalin fixed paraffin embedded sections, cells in culture, other:
    Sample preparation
    Positive control kidney, heart (weak cytoplasmic reaction) lung
    Negative control

    5) Fixation step
    Yes/No
    If yes: Fixative agent and concentration 4% buffered formalin (s above)
    Fixation time
    Fixation temperature

    6) Antigen retrieval method wather bath 98°C, EDTA 9,0, 20’

    7) Permeabilization method:
    Did you do a permeabilization step (details please) or add permeabilizing agent in any dilution buffers?
    Permeabilizing agent and concentration: none


    8) Blocking agent (eg BSA, serum…):
    Concentration Commercial kit Zytomed Polymer S Pre BLock
    Blocking time 10’
    Blocking temperature room temp

    9) Endogenous peroxidases blocked? no
    Endogenous biotins blocked? no

    10) Primary antibody (If more than one was used, describe in “additional notes”) :
    Concentration or dilution s above
    Diluent buffer Zytomed System ZUC 025
    Incubation time 60’’ room temp

    11) Secondary antibody: Zytomed System Post Block Poly AP
    Species:
    Reacts against:
    Concentration or dilution
    Diluent buffer
    Incubation time
    Fluorochrome or enzyme conjugate

    12) Washing after primary and secondary antibodies:
    Buffer ZYTOMED Systems Wash Buffer
    Number of washes

    13) Detection method AP Polymer Systems Zytomed S POLAP100

    14) How many times have you run this staining? 3
    Do you obtain the same results every time? 2nd run with different tissue (first run kidney negative)
    What steps have you altered to try and optimize the use of this antibody?

    Read More
    Answer

    Vielen Dank für Ihre Anfrage.

    Auf mein Anraten werden wir die Charge bzw den Antikörper noch einmal im Labor nachtesten, da die Färbung in der Tat kräftiger aussehen sollte, besonders bei den von Ihnen verwendeten Geweben... Ich werde Ihnen sofort mitteilen, wie die Ergebnisse geworden sind.

    In der Zwischenzeit möchte ich Sie um etwas Geduld bitten.

    Read More

    Question

    human, canc. kidney. high concentration 1/100.

    Read More
    Answer

    Vielen Dank für Ihren Anruf.

    Es tut mir leid zu hören, dass Sie Probleme mit diesem Antikörper haben.

    /Ich habe unseren Fragebogen als Word-Dokument an diese E-Mail angehängt. Durch das Ausfüllen des Fragebogens erhalten wir alle nötigen Informationen über Ihre Proben und Ihr Protokoll. Sobald Sie dieses Formular an uns zurückgeschickt haben, werden wir uns Ihr Protokoll ansehen und möglichst Veränderungsvorschläge machen, die Ihre Ergebnisse verbessern werden. Falls sich herausstellt, dass der Antikörper nicht so funktioniert, wie auf dem Datenblatt beschrieben und er innerhalb der letzten 180 Tage gekauft wurde, werden wir Ihnen gerne einen Ersatz oder eine Gutschrift schicken.



    Ich freue mich, bald wieder von Ihnen zu hören.

    Read More

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