Product Details

Uniprot No.

Target Names

TIMELESS

Alternative Names

FLJ12640 antibody; FLJ20714 antibody; hTIM antibody; Protein timeless homolog antibody; TIM antibody; TIM_HUMAN antibody; TIM1 antibody; Timeless antibody; timeless circadian clock 1 antibody; timeless circadian clock antibody; timeless homolog antibody; TIMELESS1 antibody; Tof1 homolog antibody

Species Reactivity

Human

Immunogen

A synthesized peptide from human TIMELESS protein

Immunogen Species

Homo sapiens (Human)

Conjugate

Non-conjugated

Clonality

Monoclonal

Isotype

Rabbit IgG

Clone No.

25E11

Purification Method

Affinity-chromatography

Concentration

It differs from different batches. Please contact us to confirm it.

Buffer

Rabbit IgG in 10mM phosphate buffered saline , pH 7.4, 150mM sodium chloride, 0.05% BSA, 0.02% sodium azide and 50% glycerol.

Form

Liquid

Tested Applications

ELISA, WB, IHC, FC

Recommended Dilution

Application	Recommended Dilution
WB	1:500-1:5000
IHC	1:50-1:200
FC	1:50-1:200

Protocols

ELISA Protocol
Western Blotting(WB) Protocol
Immunohistochemistry (IHC) Protocol
Flow Cytometry (FC) Protocol

Troubleshooting and FAQs

Antibody FAQs

Storage

Upon receipt, store at -20°C or -80°C. Avoid repeated freeze.

Lead Time

Basically, we can dispatch the products out in 1-3 working days after receiving your orders. Delivery time maybe differs from different purchasing way or location, please kindly consult your local distributors for specific delivery time.

Description

TIMELESS plays a central role in maintaining genome stability and regulating circadian clock function, making it an essential target for researchers investigating DNA replication, cell cycle checkpoint responses, and the molecular links between circadian rhythm disruption and cancer progression. As a core component of the replication fork protection complex, TIMELESS coordinates DNA synthesis with checkpoint signaling, positioning it at the intersection of fundamental cellular processes that are frequently dysregulated in disease states.

This recombinant monoclonal antibody, generated against a synthetic peptide from human TIMELESS protein, offers the reproducibility and sequence-defined specificity that demanding experimental workflows require. The recombinant format ensures consistent performance across experiments and eliminates the lot-to-lot variability that can complicate long-term studies or multi-site collaborations.

Validation across multiple applications demonstrates genuine experimental flexibility. Western blot analysis detects TIMELESS in MCF7 breast cancer, Jurkat T-cell leukemia, and THP-1 monocytic cell lysates, with an observed band size of approximately 150 kDa. The difference from the predicted 139 kDa molecular weight likely reflects post-translational modifications such as glycosylation or phosphorylation, which are consistent with TIMELESS biology. Immunohistochemistry staining has been optimized using human pancreatic cancer tissue sections, providing researchers studying tumor biology with a validated detection system. Flow cytometry applications enable quantitative analysis of TIMELESS expression at the single-cell level, as demonstrated in HeLa cells with clear separation from isotype control.

Whether investigating circadian clock mechanisms, DNA damage responses, or the epigenetic landscape of cancer cells, this antibody provides a reliable detection tool backed by validated performance data across complementary experimental platforms.

Usage

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

Target Background

Function

Plays an important role in the control of DNA replication, maintenance of replication fork stability, maintenance of genome stability throughout normal DNA replication, DNA repair and in the regulation of the circadian clock. Required to stabilize replication forks during DNA replication by forming a complex with TIPIN: this complex regulates DNA replication processes under both normal and stress conditions, stabilizes replication forks and influences both CHEK1 phosphorylation and the intra-S phase checkpoint in response to genotoxic stress. TIMELESS promotes TIPIN nuclear localization. Involved in cell survival after DNA damage or replication stress by promoting DNA repair. In response to double-strand breaks (DSBs), accumulates at DNA damage sites and promotes homologous recombination repair via its interaction with PARP1. May be specifically required for the ATR-CHEK1 pathway in the replication checkpoint induced by hydroxyurea or ultraviolet light. Involved in the determination of period length and in the DNA damage-dependent phase advancing of the circadian clock. Negatively regulates CLOCK|NPAS2-ARTNL/BMAL1|ARTNL2/BMAL2-induced transactivation of PER1 possibly via translocation of PER1 into the nucleus. May also play an important role in epithelial cell morphogenesis and formation of branching tubules.

Gene References into Functions

we suggest that disturbances in timeless expression may result in the disruption of the control of normal circadian rhythm, thus benefiting the survival of glioma cells and promoting carcinogenesis. PMID: 30249891
TIM rs2291738 was associated with chronotype dimensions PMID: 28708003
Inhibition of MYC significantly blocked the effects of TIM on CSC population, cell invasion and anchor-independent cell growth. TIM plays an important role in promoting breast cancer progression and may represent a novel therapeutic target for breast cancer. PMID: 28464854
Stable ectopic overexpression of TIMELESS in nasopharyngeal carcinoma cell lines conferred resistance to cisplatin-induced apoptosis in vitro and in vivo, promoted an epithelial-to-mesenchymal transition phenotype, and activated the Wnt/beta-catenin pathway and downstream gene transcription. PMID: 28583847
the 1.85 A crystal structure of a large N-terminal segment of human Timeless, spanning amino acids 1-463, is presented and this region of human Timeless harbours a partial binding site for Tipin. PMID: 28334766
results provide the first evidence that TIM is required for the correct chromatin association of the CMG complex to allow efficient DNA replication. PMID: 27587400
Our results show that TIMELESS overexpression correlates with pelvic lymph node metastasis, lymphovascular space involvement, as well as unfavorable OS and DFS in human cervical cancer. Therefore, TIMELESS expression may be a potential prognostic biomarker for cervical cancer patients PMID: 27909716
TIMELESS mutants unable to bind PARP1. TIMELESS silencing significantly impairs DNA double-strand break repair. PMID: 26456830
Data reports the crystal structure of Timeless-PARP-1 complex and provides evidence that Timeless is recruited to sites of DNA damage through PARP-1 to mediate homologous recombination repair of DNA double-strand breaks. PMID: 26344098
overexpression of TIM exerts oncogenic function in human HCCs, which is mediated via CHEK2 and EEF1A2. PMID: 25405317
TIMELESS and RORA genes may confer susceptibility to bipolar disorders and impact on circadian phenotypes PMID: 24716566
The results of this study suggest that the TIMELESS gene may be associated with the lithium prophylactic response in bipolar illness. PMID: 24636202
TIMELESS is frequently overexpressed in various types of tumor tissues, and elevated TIMELESS expression is associated with advanced tumor stage and poorer breast cancer prognosis. PMID: 24161199
Data indicate that RNAi-mediated knockdown of TIMELESS (TIM) in NIH3T3 and U2OS cells shortens the period by 1 hour and diminishes DNA damage-dependent phase advancing. PMID: 23418588
TIMELESS has a distinct contribution to suppression of chromosomal instability that is independent of its heterodimeric partner, TIPIN. PMID: 23255133
Tim-Tipin complex (or Tim alone) is able to associate with DNA polymerase epsilon bound to a 40-/80-mer DNA ligand. PMID: 23511638
All lung cancer specimens but no matched normal lung tissues were positive for TIM expression. PMID: 23173913
Kaposi's Sarcoma-associated herpesvirus episome maintenance requires Tim-assisted replication fork protection at the viral terminal repeats. PMID: 23325691
observed a significant association between stage II, III, and IV breast cancers and TIMELESS promoter hypomethylation in peripheral blood lymphocytes in 80 breast cancer cases and 80 age-matched controls PMID: 22006848
Timeless functions together with TRF1 to prevent fork collapse at telomere repeat DNA and ensure stable maintenance of telomere length and integrity. PMID: 22672906
Timeless has a function in SCC that is independent of the Tim-Tipin complex, even though the abundance of Timeless is reduced when Tipin is targeted for depletion. PMID: 21508667
Tim coordinates mitotic kinase activation with termination of DNA replication. PMID: 21573113
These findings demonstrate that Tim is essential for sustaining the episomal forms of EBV DNA in latently infected cells. PMID: 21490103
the interaction between dPERIOD and dCLOCK is TIM-dependent and modulated by light, revealing a novel and unanticipated in vivo role for TIM in circadian transcription PMID: 20980603
Data show significant association between TIMELESS variants and depression with fatigue in females, and association to depression with early morning awakening in males. PMID: 20174623
The results suggest that Timeless-Tipin functions as a replication fork stabilizer that couples DNA replication with sister chromatid cohesion established at replication forks. PMID: 20124417
TIMELESS is required for ATM-dependent CHK2 activation and G2/M checkpoint control PMID: 19996108
Down-regulation of Timeless in human cells seriously compromises replication and intra-S checkpoints, indicating an intimate connection between the circadian cycle and the DNA damage checkpoints that is in part mediated by the Timeless protein. PMID: 15798197
Tipin is a checkpoint mediator that cooperates with Tim and may regulate the nuclear relocation of Claspin in response to replication checkpoint PMID: 17102137
observation explains the similar checkpoint phenotypes observed in both Tipin- and Timeless-depleted cells PMID: 17116885
TIM and Tipin are functional orthologs of their replisome-associated yeast counterparts capable of coordinating replication with genotoxic stress responses, and distinguishes mammalian TIM from the circadian-specific paralogs. PMID: 17141802
These findings indicate that the Tim-Tipin complex mediates the UV-induced intra-S checkpoint, Tim is needed to maintain DNA replication fork movement, Tipin interacts with RPA on DNA. PMID: 17296725
HRPAP20 and TIMELESS as promising markers of tamoxifen resistance in women with ER alpha-positive breast tumors. PMID: 17909269

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Subcellular Location

Nucleus. Chromosome.

Protein Families

Timeless family

Tissue Specificity

Expressed in all tissues examined including brain, heart, lung, liver, skeletal muscle, kidney, placenta, pancreas, spleen, thymus and testis. Highest levels of expression in placenta, pancreas, thymus and testis.

Database Links

HGNC: 11813

OMIM: 603887

KEGG: hsa:8914

STRING: 9606.ENSP00000450607

UniGene: Hs.118631

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Code	CSB-RA031794A0HU
Size	US$210
	Order now
Image	Western Blot , Positive WB detected in:MCF7 whole cell lysate(30µg), Jurkat whole cell lysate(30µg), THP-1 whole cell lysate(30µg) All lanes: Timeless antibody at 1:1000 Secondary Goat polyclonal to rabbit IgG at 1/40000 dilution Predicted band size: 139 kDa Observed band size: 150 kDa Exposure time：120s IHC image of CSB-RA031794A0HU diluted at 1:100 and staining in paraffin-embedded human pancreatic cancer performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a Goat anti-rabbit polymer IgG labeled by HRP and visualized using 0.05% DAB. Overlay Peak curve showing Hela cells stained with CSB-RA031794A0HU (red line) at 1:100. The cells were fixed in 4% formaldehyde and permeated by 0.2% TritonX-100 for 10min. Then 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (1ug/110⁶cells) for 45min at 4℃. The secondary antibody used was FITC-conjugated goat anti-rabbit IgG (H+L) at 1/200 dilution for 35min at 4℃.Control antibody (green line) was Rabbit IgG (1ug/110⁶cells) used under the same conditions. Acquisition of >10, 000 events was performed.
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TIMELESS Recombinant Monoclonal Antibody

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Target Background