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    products/primary-antibodies/mtap-antibody-epr6893-ab126770.pdf

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Signal Transduction Metabolism Amino Acids
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-MTAP antibody [EPR6893] (ab126770)

  • Datasheet
  • SDS
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Western blot - Anti-MTAP antibody [EPR6893] (ab126770)
  • Western blot - Anti-MTAP antibody [EPR6893] (ab126770)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTAP antibody [EPR6893] (ab126770)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTAP antibody [EPR6893] (ab126770)
  • Immunocytochemistry/ Immunofluorescence - Anti-MTAP antibody [EPR6893] (ab126770)
  • Immunoprecipitation - Anti-MTAP antibody [EPR6893] (ab126770)
  • Flow Cytometry (Intracellular) - Anti-MTAP antibody [EPR6893] (ab126770)
  • Western blot - Anti-MTAP antibody [EPR6893] (ab126770)
  • Western blot - Anti-MTAP antibody [EPR6893] (ab126770)
  • Flow Cytometry (Intracellular) - Anti-MTAP antibody [EPR6893] (ab126770)
  • OI-RD Scanning - Anti-MTAP antibody [EPR6893] (ab126770)
  • Anti-MTAP antibody [EPR6893] (ab126770)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR6893] to MTAP
  • Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Carrier Free PE

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Protein
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Recombinant Human MTAP protein (ab132581)
Secondary
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Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
Primary
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Anti-PRAME antibody [EPR20330] (ab219650)

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Overview

  • Product name

    Anti-MTAP antibody [EPR6893]
    See all MTAP primary antibodies
  • Description

    Rabbit monoclonal [EPR6893] to MTAP
  • Host species

    Rabbit
  • Specificity

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

    Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human MTAP aa 200-300. The exact sequence is proprietary.
    Database link: Q13126

  • Positive control

    • WB: HeLa, 293T, HT29, C6, RAW 264.7, and NIH 3T3 cell lysates. ICC/IF: HeLa cells. Flow Cyt (intra): HeLa cells. IHC-P: Human kidney, mouse kidney, and human lung carcinoma tissue.
  • General notes

    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 production
    For more information see here.

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Dissociation constant (KD)

    KD = 2.60 x 10 -11 M
    Learn more about KD
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR6893
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Metabolism
    • Amino Acids
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Amino acid metabolism
    • Metabolism
    • Types of disease
    • Cancer

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-MTAP antibody [EPR6893] (ab225148)
    • PE Anti-MTAP antibody [EPR6893] (ab225149)
    • Anti-MTAP antibody [EPR6893] - BSA and Azide free (ab232417)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human MTAP knockout HeLa cell line (ab265272)
  • KO cell lysates

    • Human MTAP knockout HeLa cell lysate (ab257194)
  • Recombinant Protein

    • Recombinant Human MTAP protein (ab132581)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab126770 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt (Intra)
1/90.

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

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

WB
1/1000 - 1/10000. Detects a band of approximately 29 kDa (predicted molecular weight: 31 kDa).
IP
1/10 - 1/100.
IHC-P (1)
1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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.

ICC/IF
1/50 - 1/250.
Notes
Flow Cyt (Intra)
1/90.

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

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

WB
1/1000 - 1/10000. Detects a band of approximately 29 kDa (predicted molecular weight: 31 kDa).
IP
1/10 - 1/100.
IHC-P
1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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.

ICC/IF
1/50 - 1/250.

Target

  • Function

    Plays a major role in polyamine metabolism and is important for the salvage of both adenine and methionine.
  • Tissue specificity

    Ubiquitously expressed.
  • Sequence similarities

    Belongs to the PNP/MTAP phosphorylase family.
  • Cellular localization

    Cytoplasm.
  • Target information above from: UniProt accession Q13126 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 4507 Human
    • Entrez Gene: 66902 Mouse
    • Entrez Gene: 298227 Rat
    • Omim: 156540 Human
    • SwissProt: Q13126 Human
    • SwissProt: Q9CQ65 Mouse
    • Unigene: 193268 Human
    • Unigene: 28500 Mouse
    • Alternative names

      • 5' methylthioadenosine phosphorylase antibody
      • 5''-methylthioadenosine phosphorylase antibody
      • BDMF antibody
      • c86fus antibody
      • DMSFH antibody
      • DMSMFH antibody
      • Epididymis luminal protein 249 antibody
      • HEL 249 antibody
      • LGMBF antibody
      • MeSAdo phosphorylase antibody
      • Methylthioadenosine phosphorylase antibody
      • MSAP antibody
      • MTA phosphorylase antibody
      • MTAP antibody
      • MTAP_HUMAN antibody
      • MTAPase antibody
      • S methyl 5 thioadenosine phosphorylase antibody
      • S methyl 5' thioadenosine phosphorylase antibody
      • S-methyl-5''-thioadenosine phosphorylase antibody
      see all

    Images

    • Western blot - Anti-MTAP antibody [EPR6893] (ab126770)
      Western blot - Anti-MTAP antibody [EPR6893] (ab126770)
      All lanes : Anti-MTAP antibody [EPR6893] (ab126770) at 1/1000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : MTAP knockout HeLa cell lysate
      Lane 3 : HT-29 cell lysate
      Lane 4 : A549 cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 31 kDa
      Observed band size: 32 kDa why is the actual band size different from the predicted?



      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.

       

    • Western blot - Anti-MTAP antibody [EPR6893] (ab126770)
      Western blot - Anti-MTAP antibody [EPR6893] (ab126770)
      All lanes : Anti-MTAP antibody [EPR6893] (ab126770) at 1/10000 dilution (purified)

      Lane 1 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates
      Lane 2 : C6 (Rat glial tumor glial cell) whole cell lysates
      Lane 3 : Mouse kidney lysates

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 31 kDa



      Blocking and diluting buffer : 5% NFDM/TBST
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTAP antibody [EPR6893] (ab126770)
      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)
      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)
      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.

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

    • Flow Cytometry (Intracellular) - Anti-MTAP antibody [EPR6893] (ab126770)
      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)
      Western blot - Anti-MTAP antibody [EPR6893] (ab126770)
      All lanes : Anti-MTAP antibody [EPR6893] (ab126770) at 1/2000 dilution (purified)

      Lane 1 : HT-29 (Human colorectal adenocarcinoma epithelial cell) whole cell lysates
      Lane 2 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 31 kDa



      Blocking and diluting buffer : 5% NFDM/TBST
    • Western blot - Anti-MTAP antibody [EPR6893] (ab126770)
      Western blot - Anti-MTAP antibody [EPR6893] (ab126770)
      All lanes : Anti-MTAP antibody [EPR6893] (ab126770) at 1/1000 dilution (unpurified)

      Lane 1 : 293T cell lysates
      Lane 2 : HT29 cell lysates
      Lane 3 : C6 cell lysates
      Lane 4 : RAW 264.7 cell lysates
      Lane 5 : NIH 3T3 cell lysates

      Lysates/proteins at 10 µg per lane.

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

      Predicted band size: 31 kDa

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

       

       

    • OI-RD Scanning - Anti-MTAP antibody [EPR6893] (ab126770)
      OI-RD Scanning - Anti-MTAP antibody [EPR6893] (ab126770)
      Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD
    • Anti-MTAP antibody [EPR6893] (ab126770)
      Anti-MTAP antibody [EPR6893] (ab126770)

    Protocols

    • Flow cytometry protocols
    • Immunoprecipitation protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (2)

    Publishing research using ab126770? Please let us know so that we can cite the reference in this datasheet.

    ab126770 has been referenced in 2 publications.

    • Driehuis E  et al. Pancreatic cancer organoids recapitulate disease and allow personalized drug screening. Proc Natl Acad Sci U S A N/A:N/A (2019). PubMed: 31818951
    • Cheng XY  et al. Deletion and downregulation of MTAP contribute to the motility of esophageal squamous carcinoma cells. Onco Targets Ther 10:5855-5862 (2017). PubMed: 29270023

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

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

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Kidney)
    Antigen retrieval step
    Enzymatic - Buffer/Enzyme Used: Ventana Protease 1
    Permeabilization
    No
    Specification
    Kidney
    Blocking step
    Ventana blocking agent as blocking agent for 4 minute(s) · Concentration: 100% · Temperature: 36°C
    Fixative
    Formaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Sep 25 2018

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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