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MGMT Monoclonal antibody

MGMT Monoclonal Antibody for WB, IF/ICC, FC (Intra), ELISA

Host / Isotype

Mouse / IgG2a

Reactivity

human and More (1)

Applications

WB, IF/ICC, FC (Intra), ELISA

Conjugate

Unconjugated

CloneNo.

1H2C9

Cat no : 67476-1-Ig

Synonyms

O-6-methylguanine-DNA-alkyltransferase, Methylated-DNA--protein-cysteine methyltransferase, EC:2.1.1.63, 6-O-methylguanine-DNA methyltransferase, 1H2C9

Validation Data Gallery

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  • Western Blot (WB) analysis of various lysates using MGMT Monoclonal antibody (67476-1-Ig)

    WB analysis using 67476-1-Ig

    Various lysates were subjected to SDS PAGE followed by western blot with 67476-1-Ig (MGMT antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

  • Western Blot (WB) analysis of various lysates using MGMT Monoclonal antibody (67476-1-Ig)

    WB analysis using 67476-1-Ig

    Various lysates were subjected to SDS PAGE followed by western blot with 67476-1-Ig (MGMT antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated GAPDH Monoclonal antibody (HRP-60004) as loading control.

  • Western Blot (WB) analysis of HeLa cells using MGMT Monoclonal antibody (67476-1-Ig)

    WB analysis of HeLa using 67476-1-Ig

    WB result of MGMT antibody (67476-1-Ig; 1:4000; incubated at room temperature for 1.5 hours) with sh-Control and sh-MGMT transfected HeLa cells.

  • Immunofluorescence (IF) / fluorescent staining of HepG2 cells using MGMT Monoclonal antibody (67476-1-Ig)

    IF Staining of HepG2 using 67476-1-Ig

    Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using MGMT antibody (67476-1-Ig, Clone: 1H2C9 ) at dilution of 1:100 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L).

  • Flow cytometry (FC) experiment of Jurkat cells using MGMT Monoclonal antibody (67476-1-Ig)

    FC experiment of Jurkat using 67476-1-Ig

    1X10^6 Jurkat cells were intracellularly stained with 0.4 ug Anti-Human MGMT (67476-1-Ig, Clone:1H2C9) and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Control Antibody. Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

Tested Applications

Positive WB detected inU2OS cells, HeLa cells, HepG2 cells, LNCaP cells, Jurkat cells, A549 cells, MCF-7 cells, MOLT-4 cells, NK-92 cells, Raji cells
Positive IF/ICC detected inHepG2 cells
Positive FC (Intra) detected inJurkat cells

Recommended dilution

ApplicationDilution
Western Blot (WB)WB : 1:5000-1:50000
Immunofluorescence (IF)/ICCIF/ICC : 1:50-1:500
Flow Cytometry (FC) (INTRA)FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension
It is recommended that this reagent should be titrated in each testing system to obtain optimal results.
Sample-dependent, Check data in validation data gallery.

Published Applications

IFSee 2 publications below

Product Information

67476-1-Ig targets MGMT in WB, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human samples.

Tested Reactivity human
Cited Reactivityhuman, mouse
Host / Isotype Mouse / IgG2a
Class Monoclonal
Type Antibody
Immunogen MGMT fusion protein Ag29936
Full Name O-6-methylguanine-DNA methyltransferase
Calculated Molecular Weight 22 kDa
Observed Molecular Weight 22 kDa
GenBank Accession NumberBC000824
Gene Symbol MGMT
Gene ID (NCBI) 4255
RRIDAB_2882703
Conjugate Unconjugated
Form Liquid
Purification MethodProtein A purification
Storage Buffer PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
Storage ConditionsStore at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20oC storage. 20ul sizes contain 0.1% BSA.

Background Information

MGMT is the primary vehicle for cellular removal of alkyl lesions from the O-6 position of guanine and the O-4 position of thymine. While key to the maintenance of genomic integrity, MGMT also removes damage induced by alkylating chemotherapies, inhibiting the efficacy of cancer treatment [PMID:23065697].MGMT is the primary mechanism for the removal of alkylation damage from the O-6 position of guanine [PMID: 17482892]. The O-6 position of guanine is one of several positions in DNA bases to which alkyl groups are attached in SN1 alkylation reactions, and this repair has been well-characterized in mammalian cells and via MGMT homologs in bacteria and Archaea.[PMID: 10767620]

Protocols

Product Specific Protocols
WB protocol for MGMT antibody 67476-1-IgDownload protocol
IF protocol for MGMT antibody 67476-1-IgDownload protocol
FC protocol for MGMT antibody 67476-1-IgDownload protocol
Standard Protocols
Click here to view our Standard Protocols

Publications

SpeciesApplicationTitle
humanIF

Nucleic Acids Res

KDM6B promotes PARthanatos via suppression of O6-methylguanine DNA methyltransferase repair and sustained checkpoint response.

Authors - Mingming Yang
View Article
mouseIF

Cancer Discov

Targeting Microglial Metabolic Rewiring Synergizes with Immune Checkpoint Blockade Therapy for Glioblastoma

Authors - Zengpanpan Ye
View Article