Recombinant Anti-Hamartin antibody [EP318Y] - BSA and Azide free (ab247297)
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 - BSA and Azide free
- Suitable for: IHC-P, WB, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Hamartin antibody [EP318Y] - BSA and Azide free
See all Hamartin primary antibodies -
Description
Rabbit monoclonal [EP318Y] to Hamartin - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, WB, Flow Cyt (Intra)more details
Unsuitable for: ICC/IF -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- IHC-P: 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
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General notes
ab247297 is the carrier-free version of ab40872.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
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.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP318Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Alexa Fluor® 488 Anti-Hamartin antibody [EP318Y] (ab223387)
- Alexa Fluor® 647 Anti-Hamartin antibody [EP318Y] (ab223388)
- PE Anti-Hamartin antibody [EP318Y] (ab303208)
- APC Anti-Hamartin antibody [EP318Y] (ab303209)
- HRP Anti-Hamartin antibody [EP318Y] (ab303210)
- Alexa Fluor® 594 Anti-Hamartin antibody [EP318Y] (ab310427)
- Alexa Fluor® 555 Anti-Hamartin antibody [EP318Y] (ab311953)
- Alexa Fluor® 568 Anti-Hamartin antibody [EP318Y] (ab312426)
- Anti-Hamartin antibody [EP318Y] (ab40872)
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab247297 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 |
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 150 kDa.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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Notes |
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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. Predicted molecular weight: 150 kDa. |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
Target
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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
modificationsPhosphorylation 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
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Database links
- Entrez Gene: 7248 Human
- Omim: 605284 Human
- SwissProt: Q92574 Human
- Unigene: 370854 Human
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Alternative names
- Hamartin antibody
- kiaa0243 antibody
- LAM antibody
see all
Images
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This data was developed using ab40872, the same antibody clone in a different buffer formulation.
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.
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Anti-Hamartin antibody [EP318Y] (ab40872) at 1/1000 dilution (Purified) + HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 µg
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 kDaWe are not sure about the nature of the extra band.
This data was developed using ab40872, the same antibody clone in a different buffer formulation.
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This data was developed using ab40872, the same antibody clone in a different buffer formulation.
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. -
This data was developed using ab40872, the same antibody clone in a different buffer formulation.Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Hamartin with Purified ab40872 at 1/20 dilution (10µg/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).
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This data was developed using ab40872, the same antibody clone in a different buffer formulation.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.
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.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab247297 has not yet been referenced specifically in any publications.