Recombinant Rat CLOCK Protein

Cat.No. : CLOCK-1459R
Product Overview : Recombinant Rat CLOCK full length or partial length protein was expressed.
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Source : Mammalian Cells
Species : Rat
Tag : His
Form : Liquid or lyophilized powder
Endotoxin : < 1.0 EU per μg of the protein as determined by the LAL method.
Purity : >80%
Notes : This item requires custom production and lead time is between 5-9 weeks. We can custom produce according to your specifications.
Storage : Store it at +4 ºC for short term. For long term storage, store it at -20 ºC~-80 ºC.
Storage Buffer : PBS buffer
Gene Name : Clock clock homolog (mouse) [ Rattus norvegicus ]
Official Symbol : CLOCK
Gene ID : 60447
mRNA Refseq : NM_021856.1
Protein Refseq : NP_068628.1
MIM :
UniProt ID : Q9WVS9

Diurnal oscillations of endogenous H 2 O 2 sustained by p66 Shc regulate circadian clocks

Journal: Nature cell biology    PubMed ID: 31768048    Data: 2020/4/27

Authors: Jian-Fei Pei, Xun-Kai Li, De-Pei Liu

Article Snippet:One microgram of recombinant CLOCK protein (Creative BioMart) was incubated with 1 mM hydrogen peroxide for 10 min at 37°C, at which point dimedone was added to a final concentration of 5 mM and the samples were incubated for an additional 1 h. After the dimedone treatment, 10 mM DTT was added, and the samples were incubated for an additional 30 min at 37°C.. Subsequently, samples were incubated with 20 mM indole-3-acetic acid (IAA) for an additional 30 min, after which trypsin digestion was performed overnight.Subsequently, samples were incubated with 20 mM indole-3-acetic acid (IAA) for an additional 30 min, after which trypsin digestion was performed overnight.

a, Environment of the C195 thiol in the CLOCK:BMAL1 crystal structure. b, Representative western blot showing the interactions between recombinant BMAL1 (amino acids 49–434) and recombinant WT- or C195S-CLOCK after treatment with or without H2O2 (10?5 mM). c,d, Scatchard analysis of the equilibrium binding of recombinant BMAL1 to recombinant WT-CLOCK (c) and C195S-CLOCK (d) treated with or without H2O2 (10?6 mM). e, Scatchard analysis of the equilibrium binding of the heterodimer of recombinant BMAL1:WT-CLOCK treated with or without H2O2 (10?6 mM) to a G-box probe. f, Representative EMSA of the heterodimer of BMAL1 and WT- or C195S-CLOCK proteins treated with or without H2O2 (10?6 mM) binding to the G-box probe. g, Relative mRNA levels of Per2, Per1, Cry2, Rev-erbα, Dbp, and Tef in WT MAFs and ClockC195S MAFs treated with or without H2O2 (50 μM) for 10 h (n = 3 independent biological samples). h-m, Relative mRNA levels of Per2 (h), Per1 (i), Cry2 (j), Rev-erbα (k), Dbp (l), and Tef (m) in WT and ClockC195S MAFs over the circadian cycle (n = 3 independent biological samples per time point). P values are shown for the comparisons of ClockC195S with WT. n, Representative baseline detrended bioluminescence recordings from synchronized Clock KO MAFs rescued by WT or C195S mutant CLOCK. P values were calculated using one-way ANOVA with a Bonferroni’s post hoc test (g) and an unpaired two-tailed Student’s t test (h-m). Data are presented as the means ± SEM. n = 3 independent experiments for b,f and n = 2 independent experiments for c-e and n = 6 independent experiments for n with similar results. Source data are provided in Statistics Source Data Figure 4. Unprocessed blots are shown in Source Data Figure 4.

a, Environment of the C195 thiol in the CLOCK:BMAL1 crystal structure. b, Representative western blot showing the interactions between recombinant BMAL1 (amino acids 49–434) and recombinant WT- or C195S-CLOCK after treatment with or without H2O2 (10?5 mM). c,d, Scatchard analysis of the equilibrium binding of recombinant BMAL1 to recombinant WT-CLOCK (c) and C195S-CLOCK (d) treated with or without H2O2 (10?6 mM). e, Scatchard analysis of the equilibrium binding of the heterodimer of recombinant BMAL1:WT-CLOCK treated with or without H2O2 (10?6 mM) to a G-box probe. f, Representative EMSA of the heterodimer of BMAL1 and WT- or C195S-CLOCK proteins treated with or without H2O2 (10?6 mM) binding to the G-box probe. g, Relative mRNA levels of Per2, Per1, Cry2, Rev-erbα, Dbp, and Tef in WT MAFs and ClockC195S MAFs treated with or without H2O2 (50 μM) for 10 h (n = 3 independent biological samples). h-m, Relative mRNA levels of Per2 (h), Per1 (i), Cry2 (j), Rev-erbα (k), Dbp (l), and Tef (m) in WT and ClockC195S MAFs over the circadian cycle (n = 3 independent biological samples per time point). P values are shown for the comparisons of ClockC195S with WT. n, Representative baseline detrended bioluminescence recordings from synchronized Clock KO MAFs rescued by WT or C195S mutant CLOCK. P values were calculated using one-way ANOVA with a Bonferroni’s post hoc test (g) and an unpaired two-tailed Student’s t test (h-m). Data are presented as the means ± SEM. n = 3 independent experiments for b,f and n = 2 independent experiments for c-e and n = 6 independent experiments for n with similar results. Source data are provided in Statistics Source Data Figure 4. Unprocessed blots are shown in Source Data Figure 4.

For Research Use Only. Not intended for any clinical use. No products from Creative BioMart may be resold, modified for resale or used to manufacture commercial products without prior written approval from Creative BioMart.

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