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Rabbit Recombinant Monoclonal MTAP antibody. Suitable for IHC-P, IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples. Cited in 8 publications.


Images

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.
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Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PIPWBICC/IFFlow Cyt (Intra)
Human
Tested
Tested
Tested
Tested
Tested
Mouse
Expected
Expected
Tested
Expected
Expected
Rat
Expected
Expected
Tested
Expected
Expected

Tested
Tested

Species

Human

Dilution info

1/1000

Notes

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat

For unpurified use at 1:50 - 1:100.

Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

1/10 - 1/100

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/1000 - 1/10000

Notes

-

Species

Rat

Dilution info

1/1000 - 1/10000

Notes

-

Species

Human

Dilution info

1/1000 - 1/10000

Notes

-

Tested
Tested

Species

Human

Dilution info

1/50 - 1/250

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

1/90

Notes

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

For unpurified use at 1/100 - 1/500.

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Target data

Function

Catalyzes the reversible phosphorylation of S-methyl-5'-thioadenosine (MTA) to adenine and 5-methylthioribose-1-phosphate. Involved in the breakdown of MTA, a major by-product of polyamine biosynthesis. Responsible for the first step in the methionine salvage pathway after MTA has been generated from S-adenosylmethionine. Has broad substrate specificity with 6-aminopurine nucleosides as preferred substrates.

Alternative names

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Rabbit Recombinant Monoclonal MTAP antibody. Suitable for IHC-P, IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples. Cited in 8 publications.

Alternative names

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EPR6893

Purification technique

Affinity purification Protein A

Specificity

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

Dissociation constant

2.6 x 10-11 M

Concentration
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Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

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.

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.

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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

11 product images

  • Western blot - Anti-MTAP antibody [EPR6893] (ab126770), expandable thumbnail

    Western blot - Anti-MTAP antibody [EPR6893] (ab126770)

    anes 1- 4: Merged signal (red and green). Green - ab126770 observed at 32 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

    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 ab265272 (knockout cell lysate ab257194) was used. Wild-type and MTAP knockout samples were subjected to SDS-PAGE. ab126770 and 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 (ab216773) and 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 [EPR6893] (AB126770) 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 - Anti-MTAP antibody [EPR6893] (ab126770), expandable thumbnail

    Western blot - Anti-MTAP antibody [EPR6893] (ab126770)

    Blocking and diluting buffer : 5% NFDM/TBST

    All lanes: Western blot - Anti-MTAP antibody [EPR6893] (AB126770) at 1/10000 dilution

    Lane 1: NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates at 15 µg

    Lane 2: C6 (Rat glial tumor glial cell) whole cell lysates at 15 µg

    Lane 3: Mouse kidney lysates at 15 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051) at 1/20000 dilution

    Predicted band size: 31 kDa

  • Immunoprecipitation - Anti-MTAP antibody [EPR6893] (ab126770), expandable thumbnail

    Immunoprecipitation - Anti-MTAP antibody [EPR6893] (ab126770)

    ab126770 (purified) at 1:50 dilution (2μg) immunoprecipitating MTAP in HT-29 whole cell lysate.
    Lane 1 (input): HT-29 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate 10ug
    Lane 2 (+): ab126770 & HT-29 whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab126770 in HT-29 whole cell lysate
    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.

    All lanes: Immunoprecipitation - Anti-MTAP antibody [EPR6893] (AB126770)

    Predicted band size: 31 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTAP antibody [EPR6893] (ab126770), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTAP antibody [EPR6893] (ab126770)

    Formalin-fixed, paraffin-embedded human kidney tissue stained for MTAP with unpurified ab126770 (1/50 dilution) in immunohistochemical analysis.

    Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTAP antibody [EPR6893] (ab126770), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTAP antibody [EPR6893] (ab126770)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human lung carcinoma tissue sections labeling MTAP with Purified ab126770 at 1:1000 dilution (0.89 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

  • Immunocytochemistry/ Immunofluorescence - Anti-MTAP antibody [EPR6893] (ab126770), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-MTAP antibody [EPR6893] (ab126770)

    Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labeling MTAP with Purified ab126770 at 1/250 dilution. Cells were fixed with 4% PFA and permeabilized with 0.1% tritonX-100. ab150077 Goat anti rabbit IgG (Alexa Fluor® 488) at 1/1000 was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

  • Flow Cytometry (Intracellular) - Anti-MTAP antibody [EPR6893] (ab126770), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-MTAP antibody [EPR6893] (ab126770)

    Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling MTAP with purified ab126770 at 1/90 dilution (10 μg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

  • Western blot - Anti-MTAP antibody [EPR6893] (ab126770), expandable thumbnail

    Western blot - Anti-MTAP antibody [EPR6893] (ab126770)

    Blocking and diluting buffer : 5% NFDM/TBST

    All lanes: Western blot - Anti-MTAP antibody [EPR6893] (AB126770) at 1/2000 dilution

    Lane 1: HT-29 (Human colorectal adenocarcinoma epithelial cell) whole cell lysates at 20 µg

    Lane 2: HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051) at 1/20000 dilution

    Predicted band size: 31 kDa, 71 kDa

  • Flow Cytometry (Intracellular) - Anti-MTAP antibody [EPR6893] (ab126770), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-MTAP antibody [EPR6893] (ab126770)

    Overlay histogram showing HeLa cells stained with unpurified ab126770 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab126770, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

  • Western blot - Anti-MTAP antibody [EPR6893] (ab126770), expandable thumbnail

    Western blot - Anti-MTAP antibody [EPR6893] (ab126770)

    All lanes: Western blot - Anti-MTAP antibody [EPR6893] (AB126770) at 1/1000 dilution

    Lane 1: 293T cell lysates at 10 µg

    Lane 2: HT29 cell lysates at 10 µg

    Lane 3: C6 cell lysates at 10 µg

    Lane 4: RAW 264.7 cell lysates at 10 µg

    Lane 5: NIH 3T3 cell lysates at 10 µg

    Secondary

    All lanes: Goat anti-Rabbit HRP at 1/2000 dilution

    Predicted band size: 31 kDa

  • OI-RD Scanning - Anti-MTAP antibody [EPR6893] (ab126770), expandable thumbnail

    OI-RD Scanning - Anti-MTAP antibody [EPR6893] (ab126770)

    We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
    Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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