Recombinant Rat CLOCK Protein

• Cat.No.: CLOCK-1459R
• Product Overview: Recombinant Rat CLOCK full length or partial length protein was expressed.

Specification

• Species: Rat
• Source: Mammalian Cells
• 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 Information

• Gene Name: Clock clock homolog (mouse) [ Rattus norvegicus ]
• Official Symbol: CLOCK
• Gene ID: 60447
• mRNA Refseq: NM_021856.1
• Protein Refseq: NP_068628.1
• UniProt ID: Q9WVS9

Citation

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.