Validation Data Gallery

Filter: AllWBIPIHCFC (Intra)

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.
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.
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.
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.
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.
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).
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).
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).
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).
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).
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).
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

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

Formulation: PBS, Azide, Glycerol

PBS, Azide, Glycerol

Conjugate: Unconjugated

Unconjugated

Size/Concentration:

-/ -

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Overview

Tested Applications
Product Information
Publications (22)
Protocols

データシート［PDF］を印刷

MSDS

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

Application	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 相同性解析による交差性が予測される生物種
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
Form	Liquid
Purification Method	Antigen affinity purification
UNIPROT ID	P16455
Storage Buffer	PBS with 0.02% sodium azide and 50% glycerol{{ptg:BufferTemp}}7.3
Storage Conditions	Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage.

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
IHC protocol for MGMT antibody 17195-1-AP	Download protocol
IP protocol for MGMT antibody 17195-1-AP	Download protocol
WB protocol for MGMT antibody 17195-1-AP	Download protocol

Standard Protocols
Click here to view our Standard Protocols

Publications

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 View Article
human	WB	Nucleic Acids Res KDM6B promotes PARthanatos via suppression of O6-methylguanine DNA methyltransferase repair and sustained checkpoint response. Authors - Mingming Yang View Article
human	WB	Clin Transl Med LncRNA BC promotes lung adenocarcinoma progression by modulating IMPAD1 alternative splicing Authors - Qi Wen Chen View Article
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 View Article
human	WB	Oncogene GBP3 promotes glioblastoma resistance to temozolomide by enhancing DNA damage repair. Authors - Hui Xu View Article
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 View Article

Target

Host

Species Reactivity

Species Reactivity

Conjugate

Conjugate

Validation Data Gallery

WB analysis using 17195-1-AP

WB analysis of HeLa using 17195-1-AP

WB analysis of Jurkat using 17195-1-AP

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

IP experiment of Jurkat using 17195-1-AP

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

IHC staining of human placenta using 17195-1-AP

IHC staining of human placenta using 17195-1-AP

IHC staining of human testis using 17195-1-AP

IHC staining of human testis using 17195-1-AP

IHC staining of human spleen using 17195-1-AP

IHC staining of human spleen using 17195-1-AP

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

FC experiment of Jurkat using 17195-1-AP

MGMT Polyclonal antibody

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

-/ -

Tested Applications

Recommended dilution

Published Applications

Product Information

Background Information

Protocols

Publications

DNA damage triggers tubular endoplasmic reticulum extension to promote apoptosis by facilitating ER-mitochondria signaling.

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

LncRNA BC promotes lung adenocarcinoma progression by modulating IMPAD1 alternative splicing

MGMT activated by Wnt pathway promotes cisplatin tolerance through inducing slow-cycling cells and nonhomologous end joining in colorectal cancer

GBP3 promotes glioblastoma resistance to temozolomide by enhancing DNA damage repair.

GSK3β palmitoylation mediated by ZDHHC4 promotes tumorigenicity of glioblastoma stem cells in temozolomide-resistant glioblastoma through the EZH2-STAT3 axis.