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 Cytmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide corresponding to C-terminal residues of human Hamartin.

  • General notes

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

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 1/20000. Predicted molecular weight: 150 kDa.
IHC-P Use at an assay dependent concentration.
Flow Cyt 1/1000 - 1/10000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

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

  • 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.

  • Ab40872 staining human Hamartin in human kidneys by immunohistochemistry using paraffin embedded tissue.
  • Overlay histogram showing HeLa cells stained with ab40872 (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 (ab40872, 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) (0.1μg/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% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
  • Anti-Hamartin antibody [EP318Y] (ab40872) at 1/20000 dilution + HeLa cell lysate

    Predicted band size: 150 kDa
    Observed band size: 150 kDa

References

This product has been referenced in:
  • Platt FM  et al. Spectrum of phosphatidylinositol 3-kinase pathway gene alterations in bladder cancer. Clin Cancer Res 15:6008-17 (2009). Read more (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). Read more (PubMed: 18397877) »
See all 2 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Question
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

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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?

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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.

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Question
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.

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