Recombinant Anti-MGMT antibody [EPR4397] (ab108630)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR4397] to MGMT
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-MGMT antibody [EPR4397]
See all MGMT primary antibodies -
Description
Rabbit monoclonal [EPR4397] to MGMT -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-Pmore details
Unsuitable for: Flow Cyt,ICC/IF or IP -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: MCF7, Jurkat, HeLa, and HT29 cell lysates IHC-P: Human clear cell carcinoma, lung cancer, and spleen tissues
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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 -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR4397 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab108630 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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IHC-P |
1/800. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Antigen retrieval is recommended. See IHC antigen retrieval protocols. For unpurified use at 1/250- 1/500. |
Notes |
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WB
1/1000 - 1/10000. Detects a band of approximately 23 kDa (predicted molecular weight: 22 kDa). |
IHC-P
1/800. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. Antigen retrieval is recommended. See IHC antigen retrieval protocols. For unpurified use at 1/250- 1/500. |
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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Anti-MGMT antibody [EPR4397] (ab108630) at 1/5000 dilution (Purified) + MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 22 kDa
Observed band size: 22 kDa -
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 - unpurified ab108630 observed at 22 kDa. Red - loading control, ab8245, observed at 37 kDa.ab108630 was shown to specifically react with MGMT when MGMT knockout samples were used. Wild-type and MGMT knockout samples were subjected to SDS-PAGE. ab108630 and ab8245 (loading control to GAPDH) 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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung cancer tissue sections labeling MGMT with purified ab108630 at 1:800 dilution (0.94 µg/ml). Heat mediated antigen retrieval was performed using Citrate buffer, pH 6.0. ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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All lanes : Anti-MGMT antibody [EPR4397] (ab108630) at 1/1000 dilution (Unpurified)
Lane 1 : MCF7 cell lysate
Lane 2 : Jurkat ell lysate
Lane 3 : HeLa cell lysate
Lane 4 : HT29 cell lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 22 kDa
Observed band size: 23 kDa why is the actual band size different from the predicted? -
Unpurified ab108630, at a 1/250 dilution, staining MGMT in paraffin embedded Human clear cell carcinoma tissue by Immunohistochemistry.
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Unpurified ab108630, at a 1/250 dilution, staining MGMT in paraffin embedded Human spleen tissue by Immunohistochemistry.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (12)
ab108630 has been referenced in 12 publications.
- Zhang Q et al. miR-145-5p inhibits the proliferation of glioma stem cells by targeting translationally controlled tumor protein. J Cancer 13:1490-1500 (2022). PubMed: 35371310
- Geng X et al. Exosomal circWDR62 promotes temozolomide resistance and malignant progression through regulation of the miR-370-3p/MGMT axis in glioma. Cell Death Dis 13:596 (2022). PubMed: 35817771
- Feng SW et al. Exploring the Mechanism of Adjuvant Treatment of Glioblastoma Using Temozolomide and Metformin. Int J Mol Sci 23:N/A (2022). PubMed: 35897747
- Qiu R et al. Ginsenosides Rg1 and CK Control Temozolomide Resistance in Glioblastoma Cells by Modulating Cholesterol Efflux and Lipid Raft Distribution. Evid Based Complement Alternat Med 2022:1897508 (2022). PubMed: 36276866
- Wu K et al. LncRNA POU3F3 Contributes to Dacarbazine Resistance of Human Melanoma Through the MiR-650/MGMT Axis. Front Oncol 11:643613 (2021). PubMed: 33816296