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.
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
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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) - 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.
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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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Datasheet download
References (20)
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