MGMT Polyclonal antibody

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  WB analysis using 17195-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 17195-1-AP (MGMT antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

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  WB analysis of HeLa using 17195-1-AP

  WB result of MGMT antibody (17195-1-AP; 1:4000; incubated at room temperature for 1.5 hours) with sh-Control and sh-MGMT transfected HeLa cells.

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  WB analysis of Jurkat using 17195-1-AP

  Jurkat cells were subjected to SDS PAGE followed by western blot with 17195-1-AP (MGMT antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

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  WB analysis of MCF-7 using 17195-1-AP

  MCF7 cells were subjected to SDS PAGE followed by western blot with 17195-1-AP (MGMT antibody) at dilution of 1:800 incubated at room temperature for 1.5 hours.

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  IP experiment of Jurkat using 17195-1-AP

  IP result of anti-MGMT (IP:17195-1-AP, 4ug; Detection:17195-1-AP 1:500) with Jurkat cells lysate 3200ug.

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  IHC staining of human liver cancer using 17195-1-AP

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 17195-1-AP (MGMT antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human placenta using 17195-1-AP

  Immunohistochemical analysis of paraffin-embedded human placenta using 17195-1-AP (MGMT antibody) at dilution of 1:100 (under 10x lens).

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  IHC staining of human placenta using 17195-1-AP

  Immunohistochemical analysis of paraffin-embedded human placenta using 17195-1-AP (MGMT antibody) at dilution of 1:100 (under 40x lens).

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  IHC staining of human testis using 17195-1-AP

  Immunohistochemical analysis of paraffin-embedded human testis using 17195-1-AP (MGMT antibody) at dilution of 1:100 (under 10x lens).

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  IHC staining of human testis using 17195-1-AP

  Immunohistochemical analysis of paraffin-embedded human testis using 17195-1-AP (MGMT antibody) at dilution of 1:100 (under 40x lens).

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  IHC staining of human spleen using 17195-1-AP

  Immunohistochemical analysis of paraffin-embedded human spleen using 17195-1-AP (MGMT antibody) at dilution of 1:100 (under 10x lens).

• 

  IHC staining of human spleen using 17195-1-AP

  Immunohistochemical analysis of paraffin-embedded human spleen using 17195-1-AP (MGMT antibody) at dilution of 1:100 (under 40x lens).

• 

  IHC staining of human liver cancer using 17195-1-AP

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 17195-1-AP (MGMT antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  FC experiment of Jurkat using 17195-1-AP

  1X10^6 Jurkat cells were intracellularly stained with 0.2 ug Anti-Human MGMT (17195-1-AP) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) at dilution 1:1000 (red), or 0.2 ug Control Antibody. Cells were fixed and permeabilized with Transcription Factor Staining Buffer Kit (PF00011).

• Featured Product
• KD/KO Validated

MGMT Polyclonal Antibody for WB, IHC, FC (Intra), IP, ELISA

Cat No. 17195-1-AP

Publications(22)

Host / Isotype

Rabbit / IgG

Reactivity

human and More (1)

Applications

WB, IHC, IF, FC (Intra), IP, CoIP, ELISA

Synonyms

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

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Tested Applications

• Positive WB detected in: Jurkat cells, MCF-7 cells, HeLa cells, Raji cells
• Positive IP detected in: Jurkat cells
• Positive IHC detected in: human liver cancer tissue, human placenta tissue, human spleen tissue, human testis tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive FC (Intra) detected in: Jurkat cells

Recommended dilution

• Western Blot (WB): WB : 1:1000-1:6000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.20 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

• KD/KO: See 1 publications below
• WB: See 18 publications below
• IHC: See 4 publications below
• IF: See 1 publications below
• CoIP: See 1 publications below

Product Information

17195-1-AP targets MGMT in WB, IHC, IF, FC (Intra), IP, CoIP, ELISA applications and shows reactivity with human samples.

• Tested Reactivity: human
• Cited Reactivity: human, mouse
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen:

  CatNo: Ag9996

  Product name: Recombinant human MGMT protein

  Source: e coli.-derived, PGEX-4T

  Tag: GST

  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_2143221
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity 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	WB	Cell Res; DNA damage triggers tubular endoplasmic reticulum extension to promote apoptosis by facilitating ER-mitochondria signaling.; Authors - Pengli Zheng
human	WB	Nucleic Acids Res; KDM6B promotes PARthanatos via suppression of O6-methylguanine DNA methyltransferase repair and sustained checkpoint response.; Authors - Mingming Yang
human	WB	Clin Transl Med; LncRNA BC promotes lung adenocarcinoma progression by modulating IMPAD1 alternative splicing; Authors - Qi Wen Chen
human,mouse	WB,IHC	J Pharm Anal; MGMT activated by Wnt pathway promotes cisplatin tolerance through inducing slow-cycling cells and nonhomologous end joining in colorectal cancer; Authors - Haowei Zhang ; KD Validated
human	WB	Oncogene; GBP3 promotes glioblastoma resistance to temozolomide by enhancing DNA damage repair.; Authors - Hui Xu
human	IHC	Oncogenesis; GSK3β palmitoylation mediated by ZDHHC4 promotes tumorigenicity of glioblastoma stem cells in temozolomide-resistant glioblastoma through the EZH2-STAT3 axis.; Authors - Chenggang Zhao