MGMT Recombinant monoclonal antibody

• 

  WB analysis using 86115-5-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 89933-4-RR (MGMT antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

• 

  IHC staining of human lung cancer using 86115-5-RR

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 86115-5-RR (MGMT antibody) at dilution of 1:1000 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human lung cancer using 86115-5-RR

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 86115-5-RR (MGMT antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of MCF-7 using 86115-5-RR

  Immunofluorescent analysis of (4% PFA) fixed MCF-7 cells using MGMT antibody (89933-4-RR, Clone: 243130G10 ) at dilution of 1:800 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).

MGMT Uni-rAb^® Recombinant Antibody for WB, IHC, IF/ICC, ELISA

Cat No. 86115-5-RR

Clone No.243130G10

Host / Isotype

Rabbit / IgG

Reactivity

human

Applications

WB, IHC, IF/ICC, ELISA

Synonyms

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

---

Tested Applications

• Positive WB detected in: Jurkat cells, MCF-7 cells, Raji cells, HeLa cells, HepG2 cells, LNCaP cells
• Positive IHC detected in: human lung cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: MCF-7 cells

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:10000
• Immunohistochemistry (IHC): IHC : 1:500-1:2000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:400-1:1600

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.

Product Information

86115-5-RR targets MGMT in WB, IHC, IF/ICC, ELISA applications and shows reactivity with human samples.

• Tested Reactivity: human
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• 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
• 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).