Recombinant Anti-MGMT antibody [EPR4398] (ab108989)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR4398] to MGMT
- Suitable for: WB, IP
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-MGMT antibody [EPR4398]
See all MGMT primary antibodies -
Description
Rabbit monoclonal [EPR4398] to MGMT -
Host species
Rabbit -
Tested applications
Suitable for: WB, IPmore details
Unsuitable for: ICC/IF or IHC-P -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- MCF7 and Jurkat cell lysates
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR4398 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab108989 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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WB |
1/1000 - 1/10000. Detects a band of approximately 23 kDa (predicted molecular weight: 22 kDa).
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IP |
1/10 - 1/100.
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Notes |
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WB
1/1000 - 1/10000. Detects a band of approximately 23 kDa (predicted molecular weight: 22 kDa). |
IP
1/10 - 1/100. |
Target
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Function
Involved in the cellular defense against the biological effects of O6-methylguanine (O6-MeG) in DNA. Repairs alkylated guanine in DNA by stoichiometrically transferring the alkyl group at the O-6 position to a cysteine residue in the enzyme. This is a suicide reaction: the enzyme is irreversibly inactivated. -
Sequence similarities
Belongs to the MGMT family. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 4255 Human
- Omim: 156569 Human
- SwissProt: P16455 Human
- Unigene: 501522 Human
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Alternative names
- 6 O methylguanine DNA methyltransferase antibody
- 6-O-methylguanine-DNA methyltransferase antibody
- Agat antibody
see all
Images
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: MGMT knockout HAP1 cell lysate (20 µg)
Lane 3: MCF7 cell lysate (20 µg)
Lane 4: Jurkat cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab108989 observed at 22 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab108989 was shown to specifically react with MGMT when MGMT knockout samples were used. Wild-type and MGMT knockout samples were subjected to SDS-PAGE. ab108989 and ab8245 (loading control to MGMT) were diluted 1/1000 and 1/2000 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 1/10 000 dilution for 1 h at room temperature before imaging. -
All lanes : Anti-MGMT antibody [EPR4398] (ab108989) at 1/1000 dilution (purified)
Lane 1 : MCF7 cell lysate
Lane 2 : Jurkat cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution
Predicted band size: 22 kDa
Observed band size: 23 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST -
ab108989 (purified) at 1/30 immunoprecipitating MGMT in 10 μg MCF7 (Lanes 1 and 2, observed at 23 kDa). Lane 3 - PBS. For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution. Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
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All lanes : Anti-MGMT antibody [EPR4398] (ab108989) at 1/1000 dilution (unpurified)
Lane 1 : MCF7 cell lysate
Lane 2 : Jurkat cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 22 kDa
Observed band size: 23 kDa why is the actual band size different from the predicted?
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab108989 has not yet been referenced specifically in any publications.