CLOCK ELISA Kit (Human) (OKEH08376)

Catalog No: OKEH08376

Size:96T

Human CLOCK ELISA Standard Curve

Product Info

• Predicted Species Reactivity: Homo sapiens, Human
• Application: Enzyme-linked Immunosorbent assay-Sandwich
• ELISA Kit Detection Method: Colorimetric, OD450 nm
• ELISA Kit Principle: Aviva Systems Biology CLOCK ELISA Kit (Human) (OKEH08376) is based on standard sandwich enzyme-linked immuno-sorbent assay technology. An antibody specific for CLOCK has been pre-coated onto a 96-well plate (12 x 8 Well Strips) and blocked. Standards or test samples are added to the wells, incubated and removed. A biotinylated detector antibody specific for CLOCK is added, incubated and followed by washing. Avidin-Peroxidase Conjugate is then added, incubated and unbound conjugate is washed away. An enzymatic reaction is produced through the addition of TMB substrate which is catalyzed by HRP generating a blue color product that changes yellow after adding acidic stop solution. The density of yellow coloration read by absorbance at 450 nm is quantitatively proportional to the amount of sample CLOCK captured in well.
• ELISA Kit Range: 0.156-10ng/mL
• ELISA Kit Recovery: Mean recovery when spiking into sample matrices at concentrations within the dynamic range: 109% (n = 20)
• ELISA Kit Reproducibility: Mean Intra-assay CV%: <=5.1% (n = 20)
  Mean Inter-assay CV%: <=8.6% (n = 20)
• ELISA Kit Component:
  • CLOCK Microplate: 96 Wells (12 x 8 Well strips)
  • CLOCK Lyophilized Standard: 2
  • 100X Biotinylated CLOCK Detector Antibody: 1 x 120 uL
  • 100X Avidin-HRP Conjugate: 1 x 120 uL
  • Sample Diluent: 1 x 20 mL
  • Detector Antibody Diluent: 1 x 12 mL
  • Conjugate Diluent: 1 x 12 mL
  • 25X Wash Buffer: 1 x 30 mL
  • TMB Substrate: 1 x 10 mL
  • Stop Solution: 1 x 10 mL
• Additional Information: Function: Transcriptional activator which forms a core component of the circadian clock. The circadian clock, an internal time-keeping system, regulates various physiological processes through the generation of approximately 24 hour circadian rhythms in gene expression, which are translated into rhythms in metabolism and behavior. It is derived from the Latin roots 'circa' (about) and 'diem' (day) and acts as an important regulator of a wide array of physiological functions including metabolism, sleep, body temperature, blood pressure, endocrine, immune, cardiovascular, and renal function. Consists of two major components: the central clock, residing in the suprachiasmatic nucleus (SCN) of the brain, and the peripheral clocks that are present in nearly every tissue and organ system. Both the central and peripheral clocks can be reset by environmental cues, also known as Zeitgebers (German for 'timegivers'). The predominant Zeitgeber for the central clock is light, which is sensed by retina and signals directly to the SCN. The central clock entrains the peripheral clocks through neuronal and hormonal signals, body temperature and feeding-related cues, aligning all clocks with the external light/dark cycle. Circadian rhythms allow an organism to achieve temporal homeostasis with its environment at the molecular level by regulating gene expression to create a peak of protein expression once every 24 hours to control when a particular physiological process is most active with respect to the solar day. Transcription and translation of core clock components (CLOCK, NPAS2, ARNTL/BMAL1, ARNTL2/BMAL2, PER1, PER2, PER3, CRY1 and CRY2) plays a critical role in rhythm generation, whereas delays imposed by post-translational modifications (PTMs) are important for determining the period (tau) of the rhythms (tau refers to the period of a rhythm and is the length, in time, of one complete cycle). A diurnal rhythm is synchronized with the day/night cycle, while the ultradian and infradian rhythms have a period shorter and longer than 24 hours, respectively. Disruptions in the circadian rhythms contribute to the pathology of cardiovascular diseases, cancer, metabolic syndromes and aging. A transcription/translation feedback loop (TTFL) forms the core of the molecular circadian clock mechanism. Transcription factors, CLOCK or NPAS2 and ARNTL/BMAL1 or ARNTL2/BMAL2, form the positive limb of the feedback loop, act in the form of a heterodimer and activate the transcription of core clock genes and clock-controlled genes (involved in key metabolic processes), harboring E-box elements (5'-CACGTG-3') within their promoters. The core clock genes: PER1/2/3 and CRY1/2 which are transcriptional repressors form the negative limb of the feedback loop and interact with the CLOCK|NPAS2-ARNTL/BMAL1|ARNTL2/BMAL2 heterodimer inhibiting its activity and thereby negatively regulating their own expression. This heterodimer also activates nuclear receptors NR1D1/2 and RORA/B/G, which form a second feedback loop and which activate and repress ARNTL/BMAL1 transcription, respectively. CLOCK has an intrinsic acetyltransferase activity, which enables circadian chromatin remodeling by acetylating histones and nonhistone proteins, including its own partner ARNTL/BMAL1. Regulates the circadian expression of ICAM1, VCAM1, CCL2, THPO and MPL and also acts as an enhancer of the transactivation potential of NF-kappaB. Plays an important role in the homeostatic regulation of sleep. The CLOCK-ARNTL/BMAL1 heterodimer regulates the circadian expression of SERPINE1/PAI1, VWF, B3, CCRN4L/NOC, NAMPT, DBP, MYOD1, PPARGC1A, PPARGC1B, SIRT1, GYS2, F7, NGFR, GNRHR, BHLHE40/DEC1, ATF4, MTA1, KLF10 and also genes implicated in glucose and lipid metabolism. Represses glucocorticoid receptor NR3C1/GR-induced transcriptional activity by reducing the association of NR3C1/GR to glucocorticoid response elements (GREs) via the acetylation of multiple lysine residues located in its hinge region. Promotes rhythmic chromatin opening, regulating the DNA accessibility of other transcription factors. The CLOCK-ARNTL2/BMAL2 heterodimer activates the transcription of SERPINE1/PAI1 and BHLHE40/DEC1. The preferred binding motif for the CLOCK-ARNTL/BMAL1 heterodimer is 5'-CACGTGA-3', which contains a flanking Ala residue in addition to the canonical 6-nucleotide E-box sequence (PubMed:23229515). CLOCK specifically binds to the half-site 5'-CAC-3', while ARNTL binds to the half-site 5'-GTGA-3' (PubMed:23229515). The CLOCK-ARNTL/BMAL1 heterodimer also recognizes the non-canonical E-box motifs 5'-AACGTGA-3' and 5'-CATGTGA-3' (PubMed:23229515).
  Subcellular Location: Nucleus Cytoplasm Shuffling between the cytoplasm and the nucleus is under circadian regulation and is ARNTL/BMAL1-dependent. Phosphorylated form located in the nucleus while the nonphosphorylated form found only in the cytoplasm. Sequestered to the cytoplasm in the presence of ID2 (By similarity). Localizes to sites of DNA damage in a H2AX-independent manner.
  Unigene: Hs.436975
  SMR: O15516
  String: 9606.ENSP00000308741
  KEGG: hsa:9575
  MIM: 601851
  Pfam: PF00010
• Reconstitution and Storage: Store as indicated in product manual.
• Sensitivity: 0.089ng/mL
• Specificity: Natural and recombinant Human Circadian locomoter output cycles protein kaput
• Assay Info: Assay Methodology: Quantitative Sandwich ELISA

Target Info

• Gene Symbol: CLOCK
• Gene Full Name: clock circadian regulator
• Alias Symbols: bHLHe8;circadian locomoter output cycles kaput protein;circadian locomoter output cycles protein kaput;class E basic helix-loop-helix protein 8;clock homolog;KAT13D.
• NCBI Gene Id: 9575
• Protein Name: O15516
• Description of Target: Circadian locomoter output cycles protein kaput
• Uniprot ID: https://www.uniprot.org/uniprot/O15516
• Protein Accession #: NP_001254772.1
• Nucleotide Accession #: NM_001267843.1