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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Thymidine Kinase 1/TK1 antibody [EPR3193] (ab76495)

Reviews (2)Q&A (3)References (20)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thymidine Kinase 1/TK1 antibody [EPR3193] (ab76495)
  • Western blot - Anti-Thymidine Kinase 1/TK1 antibody [EPR3193] (ab76495)
  • Flow Cytometry (Intracellular) - Anti-Thymidine Kinase 1/TK1 antibody [EPR3193] (ab76495)
  • Western blot - Anti-Thymidine Kinase 1/TK1 antibody [EPR3193] (ab76495)
  • Western blot - Anti-Thymidine Kinase 1/TK1 antibody [EPR3193] (ab76495)
  • Immunoprecipitation - Anti-Thymidine Kinase 1/TK1 antibody [EPR3193] (ab76495)
  • Anti-Thymidine Kinase 1/TK1 antibody [EPR3193] (ab76495)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3193] to Thymidine Kinase 1/TK1
  • Suitable for: Flow Cyt (Intra), WB, IP, IHC-P
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Carrier Free

Overview

  • Product name

    Anti-Thymidine Kinase 1/TK1 antibody [EPR3193]
    See all Thymidine Kinase 1/TK1 primary antibodies

  • Description

    Rabbit monoclonal [EPR3193] to Thymidine Kinase 1/TK1

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IP, IHC-Pmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

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

  • Positive control

    • WB: HEK-293T, Jurkat, HAP1, SW620, U-2 OS, and MOLT-4 cell lysates. Human tonsil tissue lysate; IP: MOLT-4 whole cell lysate; IHC-P: Human tonsil tissue; Flow Cyt (intra): HeLa cells.

  • 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.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

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: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR3193

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab76495 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)
1/200.

For unpurified use at 1/70  ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB
1/1000. Detects a band of approximately 25 kDa (predicted molecular weight: 25 kDa).

For unpurified use at 1/5000 - 1/50000
IP
1/20.
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.
Notes
Flow Cyt (Intra)
1/200.

For unpurified use at 1/70  ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB
1/1000. Detects a band of approximately 25 kDa (predicted molecular weight: 25 kDa).

For unpurified use at 1/5000 - 1/50000
IP
1/20.
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Target

  • Sequence similarities

    Belongs to the thymidine kinase family.

  • Post-translational
    modifications

    Phosphorylated on Ser-13 in mitosis.

  • Cellular localization

    Cytoplasm.
  • Information by UniProt

  • Database links

    • Alternative names

      • cytosolic antibody
      • KITH_HUMAN antibody
      • Thymidine kinase 1 antibody
      • Thymidine kinase 1 soluble antibody
      • Thymidine kinase 1 soluble isoform antibody
      • Thymidine kinase antibody
      • Thymidine kinase cytosolic antibody
      • TK 1 antibody
      • TK 2 antibody
      • TK1 antibody
      • Tk1a antibody
      • Tk1b antibody
      • TK2 antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thymidine Kinase 1/TK1 antibody [EPR3193] (ab76495)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thymidine Kinase 1/TK1 antibody [EPR3193] (ab76495)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human tonsil tissue sections labeling Thymidine Kinase 1/TK1 with purified ab76495 at 1/200 dilution (0.85 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). The immunostaining was performed on a Leica Biosystems BOND® RX instrument. 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.
    • Western blot - Anti-Thymidine Kinase 1/TK1 antibody [EPR3193] (ab76495)
      Western blot - Anti-Thymidine Kinase 1/TK1 antibody [EPR3193] (ab76495)
      All lanes : Anti-Thymidine Kinase 1/TK1 antibody [EPR3193] (ab76495) at 1/1000 dilution (Purified)

      Lane 1 : MOLT-4 (Human lymphoblastic leukemia T lymphoblast) whole cell lysate
      Lane 2 : U-2 OS (Human bone osteosarcoma epithelial cell) whole cell lysate
      Lane 3 : SW620 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate
      Lane 4 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate
      Lane 5 : Human tonsil lysate

      Secondary
      All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

      Predicted band size: 25 kDa

    • Flow Cytometry (Intracellular) - Anti-Thymidine Kinase 1/TK1 antibody [EPR3193] (ab76495)
      Flow Cytometry (Intracellular) - Anti-Thymidine Kinase 1/TK1 antibody [EPR3193] (ab76495)

      Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Thymidine Kinase 1/TK1 with Purified ab76495 at 1/200 dilution (0.1 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488 ,ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Left). Unlabeled control - /.

       

    • Western blot - Anti-Thymidine Kinase 1/TK1 antibody [EPR3193] (ab76495)
      Western blot - Anti-Thymidine Kinase 1/TK1 antibody [EPR3193] (ab76495)
      All lanes : Anti-Thymidine Kinase 1/TK1 antibody [EPR3193] (ab76495) at 1/1000 dilution

      Lane 1 : Wild-type HEK-293T cell lysate
      Lane 2 : TK1 knockout HEK-293T cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 25 kDa
      Observed band size: 25 kDa



      Lanes 1- 2: Merged signal (red and green). Green - ab76495 observed at 25 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

       ab76495 was shown to react with Thymidine Kinase 1/TK1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266230 (knockout cell lysate ab257745) was used. Wild-type HEK-293T and TK1 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab76495 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 1000 dilution and a 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 20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-Thymidine Kinase 1/TK1 antibody [EPR3193] (ab76495)
      Western blot - Anti-Thymidine Kinase 1/TK1 antibody [EPR3193] (ab76495)

      Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
      Lane 2: TK1 knockout HAP1 whole cell lysate (20 µg)
      Lane 3: Jurkat whole cell lysate (20 µg)
      Lane 4: MOLT4 whole cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab76495 observed at 25 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab76495 was shown to specifically react with TK1 in wildtype HAP1 cells. No band was observed when TK1 knockout samples were examined. Wild-type and TK1 knockout samples were subjected to SDS-PAGE.  Ab76495 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20,000 dilution for 1 hour at room temperature before imaging.

    • Immunoprecipitation - Anti-Thymidine Kinase 1/TK1 antibody [EPR3193] (ab76495)
      Immunoprecipitation - Anti-Thymidine Kinase 1/TK1 antibody [EPR3193] (ab76495)

      Purified ab76495 at 1/20 dilution (0.8µg) immunoprecipitating Thymidine Kinase 1/TK1 in MOLT-4 whole cell lysate.
      Lane 1 (input): MOLT-4 (Human lymphoblastic leukemia T lymphoblast)
      Lane 2 (+): ab76495 + MOLT-4 whole cell lysate.
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab76495 in MOLT-4 whole cell lysate.
      VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
      Blocking Buffer and concentration: 5% NFDM/TBST.
      Diluting buffer and concentration: 5% NFDM/TBST.
      Observed band size: 24 kDa

    • Anti-Thymidine Kinase 1/TK1 antibody [EPR3193] (ab76495)
      Anti-Thymidine Kinase 1/TK1 antibody [EPR3193] (ab76495)

    References (20)

    Publishing research using ab76495? Please let us know so that we can cite the reference in this datasheet.

    ab76495 has been referenced in 20 publications.

    • Suetsugu T  et al. Mechanism of acquired 5FU resistance and strategy for overcoming 5FU resistance focusing on 5FU metabolism in colon cancer cell lines. Oncol Rep 45:N/A (2021). PubMed: 33649846
    • Ma YS  et al. microRNA-320b suppresses HNF4G and IGF2BP2 expression to inhibit angiogenesis and tumor growth of lung cancer. Carcinogenesis 42:762-771 (2021). PubMed: 33758932
    • Velazquez EJ  et al. Novel monoclonal antibodies against thymidine kinase 1 and their potential use for the immunotargeting of lung, breast and colon cancer cells. Cancer Cell Int 20:127 (2020). PubMed: 32317865
    • Mori R  et al. The inhibition of thymidine phosphorylase can reverse acquired 5FU-resistance in gastric cancer cells. Gastric Cancer 22:497-505 (2019). PubMed: 30276573
    • Levin M  et al. Surmounting Cytarabine-resistance in acute myeloblastic leukemia cells and specimens with a synergistic combination of hydroxyurea and azidothymidine. Cell Death Dis 10:390 (2019). PubMed: 31101804
    View all Publications for this product

    Customer reviews and Q&As

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    1-5 of 5 Abreviews or Q&A

    Western blot abreview for Anti-Thymidine Kinase 1/TK1 antibody [EPR3193]

     Inconclusive
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (liver, spleen, brain from 13-days of age pups and)
    Gel Running Conditions
    Reduced Denaturing (12%)
    Loading amount
    50 µg
    Specification
    liver, spleen, brain from 13-days of age pups and
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    Submitted Mar 27 2018

    Flow Cytometry abreview for Anti-Thymidine Kinase 1/TK1 antibody [EPR3193]

     Good
    Abreviews
    Application
    Flow Cytometry
    Sample
    Human Cell (T cells)
    Permeabilization
    Yes - SAPONIN
    Gating Strategy
    CD4 T cells, live
    Specification
    T cells
    Fixation
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 30 2009

    Question

    Product code: 91651

    Inquiry: To whom it may concern, I am looking for an antibody againt TK-1 for paraffin embedded mouse tumour tissue and was wondering whether you had any recommendations on positive and negative control tissue. I am specifically asking as the only information about positive control tissue I found is the human tonsil which is not applicable in mice . Many thanks for your help! Regards, Kathrin

    Read More
    Answer

    Thank you for your enquiry.

    I'm sorry to confirm this antibody is not tested in IHC on paraffin sections (IHC-P). All tested species and applications covered by the 6 month guarantee are listed on the individual datasheets. These are updated as soon as we have any new information.

    The antibody has also been tested unsuccessfully in mouse, as stated on the datasheet. Therefore, regrettably this is not the most suitable antibody for your experiments.

    I can suggest one of the following two, tested in IHC-P and mouse, would be more suitable and will be covered by our 6 month guarantee in this application and species. I can recommend to review the online datasheets for further details:

    ab59271 Anti-Thymidine Kinase 1 antibody
    Tested in: IHC-P, WB, ELISA
    Rabbit polyclonal
    Reacts with:Human, Mouse
    https://www.abcam.com/Thymidine-Kinase-1TK1-antibody-ab59271.html (or use the following: https://www.abcam.com/Thymidine-Kinase-1TK1-antibody-ab59271.html).

    ab76495 2563-1 Anti-Thymidine Kinase 1 antibody [EPR3193]
    Tested in: Flow Cyt, IP, WB, ICC/IF, IHC-P
    Rabbit monoclonal
    Reacts with: Human, Mouse
    https://www.abcam.com/Thymidine-Kinase-1TK1-antibody-EPR3193-ab76495.html (or use the following: https://www.abcam.com/Thymidine-Kinase-1TK1-antibody-EPR3193-ab76495.html).

    According to the EST expression profile on the Unigen database link on the datasheet, in mouse the Tk1 protein is expressed quite highly in lymph node and several other tissues.
    http://www.ncbi.nlm.nih.gov/UniGene/ESTProfileViewer.cgi?uglist=Mm.2661

    I can therefore recommend to consider lymph node as a positive control tissue. You may wish to do some further literature searching for more information.

    Read More

    Question

    I will be grateful if you could please answer my question regarding the 2x Anti-Thymidine Kinase 1 antibody which you show on your website.
    In particular, can you please let me know what is the difference between the rabbit monoclonal Anti-Thymidine Kinase [EPR3193] (ab76495), and the rabbit monoclonal Anti-Thymidine Kinase [EPR3194] (ab91651) one.
    I wish to use the Ab for immunohistochemistry on mouse tumor slices, but am unsure what the difference between the above 2 are.
    Thank you for attending to this email. I look forward to hearing from you.

    Read More
    Answer

    Thank you for contacting Abcam.

    The main difference between ab76495 and ab91651 is that they will have different epitope recognition sites.

    Also for ab76495, we guarantee it to react with mouse and human tissue in the following applications, ICC/IF, WB, IP, IHC-P, Flow Cyt. Whereas ab91651 will react with human tissue but not mouse and rat and will work in the following applications WB, Flow Cyt, ICC/IF. Therefore ab91651 would not work for you since you wish to use the antibody on mouse sections.

    If there is anything else I can help you with, or if you need further selecting the most appropriate antibody for your experiments, please let me know.

    Read More

    Question

    Dear Tech Support Team, Kindly advise which of the following will specificaly recognize Thymidine Kinase 1 and will not cross react with Thymidine Kinase 2. ab76495 ab57757 ab91651

    Read More
    Answer

    Thank you very much for your enquiry. Based on the immunogen sequences both Thymidine Kinase 1 (TK1) antibodies ab76495 and ab91651 are specific for TK1 and will not recognise TK2: The immunogen sequence for ab76495 is a peptide near the N-terminus (amino acids a-b) of human TK1 (SwissProt P04183), whereas ab91651 has its immunogen located at the C-terminus (amino acids y-z). These sequences do not share any homology with TK2. The TK1 antibody ab57757 has not been tested for a potential cross-reactivity with TK2. However, the immunogen is the full-length human TK1 which reveals no significant sequence similarity with human TK2 (SwissProt O00142). We therefore would not expect this antibody to cross-react with TK2. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

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