MGMT Monoclonal antibody

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  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.

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  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.

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  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.

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  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 AffiniPure Goat Anti-Mouse IgG(H+L).

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  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 AffiniPure 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.

• Featured Product
• KD/KO Validated

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

Cat No. 67476-1-Ig

Clone No.1H2C9

Publications(4)

Host / Isotype

Mouse / IgG2a

Reactivity

human and More (1)

Applications

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

Synonyms

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

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Tested Applications

• Positive WB detected in: U2OS 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 in: HepG2 cells
• Positive FC (Intra) detected in: Jurkat cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunofluorescence (IF)/ICC: IF/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

• WB: See 2 publications below
• IF: See 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 Reactivity: human, mouse
• Host / Isotype: Mouse / IgG2a
• Class: Monoclonal
• Type: Antibody
• Immunogen:

  CatNo: Ag29936

  Product name: Recombinant human MGMT protein

  Source: e coli.-derived, PET28a

  Tag: 6*His

  Domain: 1-207 aa of BC000824

  Sequence: MDKDCEMKRTTLDSPLGKLELSGCEQGLHEIKLLGKGTSAADAVEVPAPAAVLGGPEPLMQCTAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLKVVKFGEVISYQQLAALAGNPKAARAVGGAMRGNPVPILIPCHRVVCSSGAVGNYSGGLAVKEWLLAHEGHRLGKPGLGGSSGLAGAWLKGAGATSGSPPAGRN

• Full Name: O-6-methylguanine-DNA methyltransferase
• Calculated Molecular Weight: 22 kDa
• Observed Molecular Weight: 22 kDa
• GenBank Accession Number: BC000824
• Gene Symbol: MGMT
• Gene ID (NCBI): 4255
• RRID: AB_2882703
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• UNIPROT ID: P16455
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC 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]

Publications

• Human
• Mouse
• Rat
• Monkey
• Pig

Species	Application	Title
human	IF	Nucleic Acids Res; KDM6B promotes PARthanatos via suppression of O6-methylguanine DNA methyltransferase repair and sustained checkpoint response.; Authors - Mingming Yang
mouse	IF	Cancer Discov; Targeting Microglial Metabolic Rewiring Synergizes with Immune Checkpoint Blockade Therapy for Glioblastoma; Authors - Zengpanpan Ye
human	WB	Nat Commun; Transcript errors generate amyloid-like proteins in huwman cells; Authors - Claire S Chung
human	WB	J Colloid Interface Sci; Brain-targeted polymersomes mediating RNAi of STAT3 sensitize glioblastoma to temozolomide and immunotherapy; Authors - Mingyu Xia