CRY1 Overexpression Lysate

Western Blot: Cryptochrome I Overexpression Lysate (Adult Normal) [NBL1-09495] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Cryptochrome I.

• Reactivity: Hu
• Applications: WB

CRY1 Overexpression Lysate Summary

• Description:

  CRY1 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC207152
  
  Accession#: NM_004075
  
  Protein Tag: C-MYC/DDK
  
  You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.
• Gene: CRY1

Applications/Dilutions

• Dilutions:
  • Western Blot
• Application Notes: This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.
  
  Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
• Theoretical MW: 66.2 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at -80C. Avoid freeze-thaw cycles.
• Buffer: RIPA buffer

Lysate Details for Array

• Type: Overexpression

Notes

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

Alternate Names for CRY1 Overexpression Lysate

• CRY1
• cryptochrome 1 (photolyase-like)
• Cryptochrome I
• PHLL1
• PHLL1cryptochrome-1
• photolyase-like cryptochrome 1

Background

Various biochemical, physiological and behavioural processes display circadian rhythms controlled by an internal biological clock. The central "gears" driving this clock appear to be composed of an autoregulatory transcription/posttranslation-based feedback loop. Cryptochrome 1 (CRY1) and 2 (CRY2) are DNA-binding flavoproteins that bear some homology to blue-light receptors and photolyases. In Drosophila, CRY is a photoreceptor for the circadian clock where it binds to the clock component TIM in a light-dependent fashion and blocks its function. Mammalian Cryptochrome 1 and Cryptochrome 2 function via light-independent interactions with circadian genes CLOCK and BMAL1, as well as with PER1, PER2, and TIM. They seem to act as light-independent components of the circadian clock and probably regulate Per1 transcriptional cycling via interactions with both the activator and its feedback inhibitors. Mutant mice not expressing the Cryptochrome 1 or Cryptochrome 2 protein display accelerated and delayed periodicity of locomotor activity, respectively. It appears that the combination of both proteins working together is essential to synchronize the organism to circadian phases. A critical balance between Cryptochrome 1 and Cryptochrome 2 is required for proper clock function; in complete darkness, double-mutant mice present with instantaneous arrhythmicity, indicating the absence of an internal circadian clock. Cryptochromes (Cry 1 and 2) are blue, ultraviolet-A photoreceptor pigment proteins that are involved circadian rhythm regulation in plants and animals. In mammals, Cry 1 and 2 expression oscillates with respect to the daily light-dark cycle in the suprachiasmatic nucleus of the hypothalamus. These proteins localize to the cell nucleus, interact with each of the Per proteins, and assist in the translocation of Per from the cytoplasm to nucleus.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Publications for Cryptochrome I Lysate (NBL1-09495)(1)

Publication using NBL1-09495

• Zeng Z, Wei J, Liu Y et al. Magnetoreception of Photoactivated Cryptochrome 1 in Electrochemistry and Electron Transfer ACS Omega. 2018-04-30 [PMID: 31458694] (Human)

Bioinformatics

• Gene Symbol: CRY1