Validated using a knockout cell line
Recombinant
RabMAb

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

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: ICC/IF, WB, IP, IHC-P, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Thymidine Kinase 1/TK1 aa 1-100. The exact sequence is proprietary.

  • Positive control

    • WB: Molt-4 cell lysate; IHC-P: human tonsil tissue.
  • General notes

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

     This product was previously labelled as Thymidine Kinase 1

     

    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 rabbit monoclonal antibody.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 9% PBS, 40% Glycerol, 0.05% BSA, 50% Tissue culture supernatant
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    EPR3193
  • Isotype

    IgG
  • Research areas

Applications

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
ICC/IF 1/100.
WB 1/5000 - 1/50000. Detects a band of approximately 25 kDa (predicted molecular weight: 25 kDa).
IP 1/10.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt 1/70.

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

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

  • 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.

  • Immunohistochemical analysis of paraffin-embedded human tonsils using 1/100 ab76495

    Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

  • Overlay histogram showing HeLa cells stained with ab76495 (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 (ab76495, 1/50 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • Anti-Thymidine Kinase 1/TK1 antibody [EPR3193] (ab76495) at 1/50000 dilution + Molt-4
    cell lysate at 10 µg

    Secondary
    HRP labelled goat anti-rabbit at 1/1000 dilution

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

References

This product has been referenced in:

  • Foloppe J  et al. The Enhanced Tumor Specificity of TG6002, an Armed Oncolytic Vaccinia Virus Deleted in Two Genes Involved in Nucleotide Metabolism. Mol Ther Oncolytics 14:1-14 (2019). Read more (PubMed: 31011628) »
  • Lopez-Gomez C  et al. Bioavailability and cytosolic kinases modulate response to deoxynucleoside therapy in TK2 deficiency. EBioMedicine 46:356-367 (2019). Read more (PubMed: 31383553) »
See all 13 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

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

Abcam user community

Verified customer

Submitted Mar 27 2018

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/index.html?datasheet=59271 (or use the following: https://www.abcam.com/index.html?datasheet=59271).

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/index.html?datasheet=76495 (or use the following: https://www.abcam.com/index.html?datasheet=76495).

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

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

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

Abcam user community

Verified customer

Submitted Oct 30 2009

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