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Catalog No: OKCD02092 (Formerly GWB-KBBKT4)
Size:96WELLS
Price: $750.00
SKU
OKCD02092
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Datasheets/ManualsPrintable datasheet for Clock ELISA Kit (Mouse) (OKCD02092)
Product Info
Predicted Species ReactivityMouse|Mus musculus
ApplicationEnzyme-linked Immunosorbent assay-Sandwich
ELISA Kit Detection MethodColorimetric
ELISA Kit Duration3h
ELISA Kit PrincipleThe test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Circadian Locomoter Output Cycles Protein Kaput (CLOCK). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Circadian Locomoter Output Cycles Protein Kaput (CLOCK). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Circadian Locomoter Output Cycles Protein Kaput (CLOCK), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm +/- 10nm. The concentration of Circadian Locomoter Output Cycles Protein Kaput (CLOCK) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
ELISA Kit Range0.156-10ng/mL
ELISA Kit ReproducibilityIntra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Circadian Locomoter Output Cycles Protein Kaput (CLOCK) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Circadian Locomoter Output Cycles Protein Kaput (CLOCK) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%
ELISA Kit Component
ComponentAmount
Anti-Clock Microplate96 Wells (12 x 8 Well Strips)
Clock Lyophilized Standard2 x 10 ng
100X Biotinylated Clock Detector Antibody120 uL
Avidin/HRP Conjugate120 uL
Standard Diluent1 x 20 mL
Detector Antibody Diluent1 x 12 mL
Conjugate Diluent1 x 12 mL
30X Wash Buffer1 x 20 mL
TMB Substrate1 x 9 mL
Stop Solution1 x 6 mL
Reconstitution and Storage2°C to 8°C|-20°C
Sample TypeTissue homogenates, cell lysates and other biological fluids
Sensitivity< 0.063 ng/mL
SpecificityThis assay has high sensitivity and excellent specificity for detection of Circadian Locomoter Output Cycles Protein Kaput (CLOCK). No significant cross-reactivity or interference between Circadian Locomoter Output Cycles Protein Kaput (CLOCK) and analogues was observed.
Assay InfoAssay Methodology: Quantitative Sandwich Immunoassay
Gene SymbolClock
Gene Full Namecircadian locomotor output cycles kaput
Alias Symbols5330400M04Rik;bHLHe;circadian locomoter output cycles protein kaput;KAT13;KAT13D.
NCBI Gene Id12753
Protein NameCircadian locomoter output cycles protein kaput
Description of TargetTranscriptional 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. 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. Promotes rhythmic chromatin opening, regulating the DNA accessibility of other transcription factors. May play a role in spermatogenesis; contributes to the chromatoid body assembly and physiology. 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 (By similarity). CLOCK specifically binds to the half-site 5'-CAC-3', while ARNTL binds to the half-site 5'-GTGA-3' (By similarity). The CLOCK-ARNTL/BMAL1 heterodimer also recognizes the non-canonical E-box motifs 5'-AACGTGA-3' and 5'-CATGTGA-3'. CLOCK has an intrinsic acetyltransferase activity, which enables circadian chromatin remodeling by acetylating histones and nonhistone proteins, including its own partner ARNTL/BMAL1. 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. The acetyltransferase activity of CLOCK is as important as its transcription activity in circadian control. Acetylates metabolic enzymes IMPDH2 and NDUFA9 in a circadian manner (By similarity). Facilitated by BMAL1, rhythmically interacts and acetylates argininosuccinate synthase 1 (ASS1) leading to enzymatic inhibition of ASS1 as well as the circadian oscillation of arginine biosynthesis and subsequent ureagenesis (PubMed:28985504).
Uniprot IDO08785
Protein Accession #NP_001276755.1
Nucleotide Accession #NM_001289826.1
Protein Size (# AA)855
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