Recombinant 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
Related conjugates and formulations
Overview
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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.
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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.
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3193 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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 |
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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. |
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WB |
1/1000. Detects a band of approximately 25 kDa (predicted molecular weight: 25 kDa).
For unpurified use at 1/5000 - 1/50000 |
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IP |
1/20.
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IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Notes |
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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. |
Target
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Sequence similarities
Belongs to the thymidine kinase family. -
Post-translational
modificationsPhosphorylated on Ser-13 in mitosis. -
Cellular localization
Cytoplasm. - Information by UniProt
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Database links
- Entrez Gene: 7083 Human
- Omim: 188300 Human
- SwissProt: P04183 Human
- Unigene: 515122 Human
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Alternative names
- cytosolic antibody
- KITH_HUMAN antibody
- Thymidine kinase 1 antibody
see all
Images
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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.
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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 -
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 - /.
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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 kDaLanes 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.
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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.
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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
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (23)
ab76495 has been referenced in 23 publications.
- Xie H et al. Assessing the Potential Prognostic and Immunological Role of TK1 in Prostate Cancer. Front Genet 13:778850 (2022). PubMed: 35559045
- Mori R et al. The mechanism underlying resistance to 5-fluorouracil and its reversal by the inhibition of thymidine phosphorylase in breast cancer cells. Oncol Lett 24:311 (2022). PubMed: 35949616
- Huang X et al. La protein regulates protein expression by binding with the mRNAs of target genes and participates the pathological process of ovarian cancer. Front Oncol 12:763480 (2022). PubMed: 36110943
- 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