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  1. Link

    ent1-antibody-sp120-ab182023.pdf

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Signal Transduction Metabolism Plasma Membrane Channels
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-ENT1 antibody [SP120] (ab182023)

  • Datasheet
  • SDS
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Western blot - Anti-ENT1 antibody [SP120] (ab182023)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ENT1 antibody [SP120] (ab182023)
  • Immunocytochemistry - Anti-ENT1 antibody [SP120] (ab182023)
  • Flow Cytometry - Anti-ENT1 antibody [SP120] (ab182023)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ENT1 antibody [SP120] (ab182023)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ENT1 antibody [SP120] (ab182023)
  • Anti-ENT1 antibody [SP120] (ab182023)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [SP120] to ENT1
  • Suitable for: Flow Cyt, IHC-P, WB, ICC
  • Knockout validated
  • Reacts with: Human

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Overview

  • Product name

    Anti-ENT1 antibody [SP120]
    See all ENT1 primary antibodies
  • Description

    Rabbit monoclonal [SP120] to ENT1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt, IHC-P, WB, ICCmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • IHC-P: Human Pancreas, Human tonsil, Human pancreas, and Human colon carcinoma tissue; Flow Cyt: HepG2 cells; ICC: HAP1 cells; WB: HAP1 cell lysate.
  • 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.

    This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.

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.
  • Storage buffer

    pH: 7.60
    Preservative: 0.1% Sodium azide
    Constituents: PBS, 1% BSA
  • Concentration information loading...
  • Purity

    Protein A/G purified
  • Purification notes

    Purified from TCS by protein A/G.
  • Clonality

    Monoclonal
  • Clone number

    SP120
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Metabolism
    • Plasma Membrane
    • Channels

Associated products

  • Alternative Versions

    • Anti-ENT1 antibody [SP120] - BSA and Azide free (ab240258)
  • Isotype control

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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab182023 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
Use at an assay dependent concentration.
IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
1/1000. Predicted molecular weight: 50 kDa.
ICC
1/200.
Notes
Flow Cyt
Use at an assay dependent concentration.
IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
1/1000. Predicted molecular weight: 50 kDa.
ICC
1/200.

Target

  • Function

    Mediates both influx and efflux of nucleosides across the membrane (equilibrative transporter). It is sensitive (ES) to low concentrations of the inhibitor nitrobenzylmercaptopurine riboside (NBMPR) and is sodium-independent. It has a higher affinity for adenosine. Inhibited by dipyridamole and dilazep (anticancer chemotherapeutics drugs).
  • Tissue specificity

    Expressed in heart, brain, mammary gland, erythrocytes and placenta, and also in fetal liver and spleen.
  • Sequence similarities

    Belongs to the SLC29A transporter family.
  • Post-translational
    modifications

    Glycosylated.
  • Cellular localization

    Basolateral cell membrane. Apical cell membrane. Predominantly localized in the basolateral membrane in polarised MDCK cells.
  • Target information above from: UniProt accession Q99808 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 2030 Human
    • Omim: 602193 Human
    • SwissProt: Q99808 Human
    • Unigene: 25450 Human
    • Alternative names

      • Equilibrative NBMPR-sensitive nucleoside transporter antibody
      • equilibrative nitrobenzylmercaptopurine riboside (NBMPR)-sensitive nucleoside transporter antibody
      • Equilibrative nitrobenzylmercaptopurine riboside-sensitive nucleoside transporter antibody
      • Equilibrative nucleoside transporter 1 antibody
      • es-type antibody
      • MGC1465 antibody
      • MGC3778 antibody
      • Nucleoside transporter antibody
      • Nucleoside transporter, es-type antibody
      • OTTHUMP00000016506 antibody
      • OTTHUMP00000016507 antibody
      • OTTHUMP00000016508 antibody
      • OTTHUMP00000016509 antibody
      • OTTHUMP00000016510 antibody
      • OTTHUMP00000016511 antibody
      • OTTHUMP00000016512 antibody
      • S29A1_HUMAN antibody
      • Slc29a1 antibody
      • solute carrier family 29 (equilibrative nucleoside transporter), member 1 antibody
      • solute carrier family 29 (nucleoside transporters), member 1 antibody
      • Solute carrier family 29 member 1 antibody
      see all

    Images

    • Western blot - Anti-ENT1 antibody [SP120] (ab182023)
      Western blot - Anti-ENT1 antibody [SP120] (ab182023)
      All lanes : Anti-ENT1 antibody [SP120] (ab182023) at 1/1000 dilution

      Lane 1 : Wild-type HAP1 cell lysate
      Lane 2 : SLC29A1 knockout HAP1 cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 50 kDa



      ab182023 was shown to react with SLC29A1 in wild-type HAP1 cells in Western blot with loss of signal observed in a SLC29A1 knockout cell line. Wild-type HAP1 and SLC29A1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab182023 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with goat anti-rabbit HRP secondary antibodies at 0.2ug/mL before imaging. These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ENT1 antibody [SP120] (ab182023)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ENT1 antibody [SP120] (ab182023)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon carcinoma tissue sections labeling ENT1 with Purified ab182023 at 1/100 dilution (0.77 µg/ml). Heat mediated antigen retrieval was performed Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    • Immunocytochemistry - Anti-ENT1 antibody [SP120] (ab182023)
      Immunocytochemistry - Anti-ENT1 antibody [SP120] (ab182023)

      ab182023 was shown to react with SLC29A1 in wild-type HAP1 cells in Immunocytochemistry with loss of signal observed in a SLC29A1 knockout cell line. Wild-type and knockout cells were mixed and pelleted at a 1:1 ratio on coverslips. The cells were fixed with 4% paraformaldehyde (15 min) then permeabilized with 0.1% Triton X-100 (10min) and then blocked with . The cells were then incubated with ab182023 at 1/200 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat anti-rabbit secondary antibody to (Alexa Fluor® 555) at 0.5 μg/ml. Acquisition of the green (wild-type), red (antibody staining) and far-red (knockout) channels was performed. Representative grayscale images of the red channel are shown. Wild-type and knockout cells are outlined with yellow and magenta dashed line, respectively. Schematic representation of the mosaic strategy used is shown on the bottom-right panel. Image was acquired with a Zeiss(LSM-880). These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

    • Flow Cytometry - Anti-ENT1 antibody [SP120] (ab182023)
      Flow Cytometry - Anti-ENT1 antibody [SP120] (ab182023)

      Flow Cytometry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling ENT1 with purified ab182023 at (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody. Isotype control - Rabbit monoclonal IgG (ab172730) / Black. Unlabeled control - Unlabelled cells / blue.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ENT1 antibody [SP120] (ab182023)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ENT1 antibody [SP120] (ab182023)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human pancreas tissue sections labeling ENT1 with Purified ab182023 at 1/100 dilution (0.77 µg/ml). Heat mediated antigen retrieval was performed Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ENT1 antibody [SP120] (ab182023)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ENT1 antibody [SP120] (ab182023)

      Immunohistochemical analysis of formalin-fixed paraffin-embedded Human pancreas tissue labeling ENT1 with ab182023.

    • Anti-ENT1 antibody [SP120] (ab182023)
      Anti-ENT1 antibody [SP120] (ab182023)

    Protocols

    • Immunohistochemistry protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (2)

    Publishing research using ab182023? Please let us know so that we can cite the reference in this datasheet.

    ab182023 has been referenced in 2 publications.

    • Heskamp S  et al. Response Monitoring with [18F]FLT PET and Diffusion-Weighted MRI After Cytotoxic 5-FU Treatment in an Experimental Rat Model for Colorectal Liver Metastases. Mol Imaging Biol 19:540-549 (2017). PubMed: 27798786
    • Oba A  et al. Clinical application of the biomarkers for the selection of adjuvant chemotherapy in pancreatic ductal adenocarcinoma. J Hepatobiliary Pancreat Sci 23:480-8 (2016). PubMed: 27247050

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