Recombinant Anti-MGMT antibody [EPR4398] (ab108989)
- Datasheet
- References
- Protocols
Overview
-
Product nameAnti-MGMT antibody [EPR4398]
See all MGMT primary antibodies -
DescriptionRabbit monoclonal [EPR4398] to MGMT
-
Host speciesRabbit
-
Tested applicationsSuitable for: WB, IPmore details
Unsuitable for: ICC/IF or IHC-P -
Species reactivityReacts with: Human
-
Immunogen
Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human MGMT aa 150 to the C-terminus.
-
Positive control
- MCF7 and Jurkat cell lysates
-
General notes
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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.
This product is a recombinant rabbit monoclonal antibody.
Properties
-
FormLiquid
-
Storage instructionsShipped 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 bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS -
Concentration information loading... -
PurityProtein A purified
-
ClonalityMonoclonal
-
Clone numberEPR4398
-
IsotypeIgG
-
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
Applications
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 |
|---|---|---|
| WB | 1/1000 - 1/10000. Detects a band of approximately 23 kDa (predicted molecular weight: 22 kDa). | |
| IP | 1/10 - 1/100. |
Target
-
FunctionInvolved 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 similaritiesBelongs to the MGMT family.
-
Cellular localizationNucleus.
- Information by UniProt
-
Database links
- Entrez Gene: 4255 Human
- Omim: 156569 Human
- SwissProt: P16455 Human
- Unigene: 501522 Human
-
Alternative names
- 6 O methylguanine DNA methyltransferase antibody
- 6-O-methylguanine-DNA methyltransferase antibody
- Agat antibody
see all
Images
-
![Western blot - Anti-MGMT antibody [EPR4398] (ab108989)](https://www.abcam.com/ps/products/108/ab108989/Images/ab108989-254226-anti-mgmt-antibody-epr4398-western-blot.jpg)
Western blot - Anti-MGMT antibody [EPR4398] (ab108989)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. -
![Western blot - Anti-MGMT antibody [EPR4398] (ab108989)](https://www.abcam.com/ps/products/108/ab108989/Images/ab108989-254660-anti-mgmt-antibody-epr4398-western-blot.jpg)
Western blot - Anti-MGMT antibody [EPR4398] (ab108989)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 -
![Immunoprecipitation - Anti-MGMT antibody [EPR4398] (ab108989)](https://www.abcam.com/ps/products/108/ab108989/Images/ab108989-254022-anti-mgmt-antibody-epr4398-immunoprecipitation.jpg)
Immunoprecipitation - Anti-MGMT antibody [EPR4398] (ab108989)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
-
![Western blot - Anti-MGMT antibody [EPR4398] (ab108989)](https://www.abcam.com/ps/products/108/ab108989/Images/ab108989-121517-anti-mgmt-antibody-epr4398-western-blot.jpg)
Western blot - Anti-MGMT antibody [EPR4398] (ab108989)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?
Datasheets and documents
References
ab108989 has not yet been referenced specifically in any publications.
