MTAP KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon1 and Insertion of the selection cassette in exon1.
Images
Key facts
- Cell type
HeLa
- Species or organism
Human
- Tissue
Cervix
- Knockout validation
Sanger Sequencing, Western blot
- Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon1 and Insertion of the selection cassette in exon1.
Alternative names
5' methylthioadenosine phosphorylase, 5''-methylthioadenosine phosphorylase, BDMF, DMSFH, DMSMFH, Epididymis luminal protein 249, HEL 249, LGMBF, MSAP, MTA phosphorylase, MTAP_HUMAN, MTAPase, MeSAdo phosphorylase, Methylthioadenosine phosphorylase, S methyl 5 thioadenosine phosphorylase, S methyl 5' thioadenosine phosphorylase, S-methyl-5''-thioadenosine phosphorylase, c86fus
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MTAP KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon1 and Insertion of the selection cassette in exon1.
Key facts
- Cell type
HeLa
- Species or organism
Human
- Tissue
Cervix
- Knockout validation
Sanger Sequencing, Western blot
- Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon1 and Insertion of the selection cassette in exon1.
- Disease
Adenocarcinoma
- Concentration
Properties
- Gene name
MTAP
- Gene editing type
Knockout
- Gene editing method
CRISPR technology
- Knockout validation
Sanger Sequencing, Western blot
Quality control
- STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
EU: 2 US: 2
- Adherent/suspension
Adherent
- Gender
Female
Storage
- Shipped at conditions
Ambient - Can Ship with Ice
- Appropriate short-term storage conditions
-20°C
- Appropriate long-term storage conditions
-20°C
Notes
Knockout cell lysate achieved by CRISPR/Cas9.
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
Supplementary info
This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
MTAP also known as methylthioadenosine phosphorylase is an essential enzyme that breaks down 5'-methylthioadenosine (MTA) a byproduct of polyamine synthesis. MTAP has a molecular weight of about 31 kDa. The enzyme converts MTA into adenine and 5-methylthioribose-1-phosphate which re-enter the methionine and adenine salvage pathways. MTAP is widely expressed in most tissues but its activity is especially high in the liver and kidney. Alternative names for MTAP include 2G4 and MTAP-A.
- Biological function summary
Methylthioadenosine phosphorylase plays a significant role in the salvage pathways for methionine and adenine critical for cellular growth and proliferation. MTAP operates as a part of a complex metabolic network involved in polyamine metabolism. In addition to its metabolic functions MTAP contributes to the regulation of the immune response and cell cycle. MTAP immunohistochemistry is often used to study its expression patterns in various tissues.
- Pathways
Methylthioadenosine phosphorylase participates importantly in the polyamine biosynthesis and methionine salvage pathways. These pathways are integral for maintaining cellular homeostasis and nucleotide pools. MTAP works closely with proteins such as methionine adenosyltransferase (MAT) and adenosylmethionine decarboxylase (AMD). In concert they facilitate the regeneration of methionine highlighting MTAP's role in cellular adaptation to metabolic demands.
- Associated diseases and disorders
Methylthioadenosine phosphorylase deficiency or deletion is linked to certain cancers such as gliomas and lymphomas. Loss of MTAP function is often associated with co-deletion of the tumor suppressor protein p16INK4a observed in various malignancies. This deletion can lead to an accumulation of MTA creating a toxic environment that promotes cancer cell proliferation. MTAP and its interaction with proteins like p16INK4a highlight its relevance as a potential target for therapeutic intervention in cancer treatment programs.
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5 product images
Western blot - Human MTAP knockout HeLa cell lysate (ab257194)
Lane 1: Wild-type HeLa cell lysate (20 μg)
Lane 2: MTAP knockout HeLa cell lysate (20 μg)
Lane 3: HT-29 cell lysate (20 μg)
Lane 4: A549 cell lysate (20 μg)
Lanes 1-4: Merged signal (red and green). Green - Anti-MTAP antibody [EPR22570-76] ab254265 observed at 32 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
Anti-MTAP antibody [EPR22570-76] ab254265 Anti-MTAP antibody [EPR22570-76] was shown to specifically react with MTAP in wild-type HeLa cells. Loss of signal was observed when knockout cell line Human MTAP knockout HeLa cell line ab265272 (knockout cell lysate ab257194) was used. Wild-type and MTAP knockout samples were subjected to SDS-PAGE. Anti-MTAP antibody [EPR22570-76] ab254265 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-MTAP antibody [EPR22570-76] (Anti-MTAP antibody [EPR22570-76] ab254265) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: MTAP knockout HeLa cell lysate at 20 µg
Lane 3: HT-29 cell lysate at 20 µg
Lane 4: A549 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 31 kDa
Observed band size: 32 kDa
Western blot - Human MTAP knockout HeLa cell lysate (ab257194)
Lane 1: Wild-type HeLa cell lysate (20 μg)
Lane 2: MTAP knockout HeLa cell lysate (20 μg)
Lane 3: HT-29 cell lysate (20 μg)
Lane 4: A549 cell lysate (20 μg)
anes 1- 4: Merged signal (red and green). Green - Anti-MTAP antibody [EPR6893] ab126770 observed at 32 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.
Anti-MTAP antibody [EPR6893] ab126770 Anti-MTAP antibody [EPR6893] was shown to specifically react with MTAP in wild-type HeLa cells. Loss of signal was observed when knockout cell line Human MTAP knockout HeLa cell line ab265272 (knockout cell lysate ab257194) was used. Wild-type and MTAP knockout samples were subjected to SDS-PAGE. Anti-MTAP antibody [EPR6893] ab126770 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-MTAP antibody [EPR22570-76] (Anti-MTAP antibody [EPR22570-76] ab254265) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: MTAP knockout HeLa cell lysate at 20 µg
Lane 3: HT-29 cell lysate at 20 µg
Lane 4: A549 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 31 kDa
Observed band size: 32 kDa
Sanger Sequencing - Human MTAP knockout HeLa cell lysate (ab257194)
Allele-2: Insertion of the selection cassette in exon1
Sanger Sequencing - Human MTAP knockout HeLa cell lysate (ab257194)
Allele-3: Insertion of the selection cassette in exon1
Sanger Sequencing - Human MTAP knockout HeLa cell lysate (ab257194)
Allele-1: 1 bp insertion in exon1
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