Overview

  • Product name

    Anti-Cryptochrome I/CRY1 antibody
    See all Cryptochrome I/CRY1 primary antibodies
  • Description

    Mouse monoclonal to Cryptochrome I/CRY1
  • Host species

    Mouse
  • Tested applications

    Suitable for: WB, ICC/IF, IHC-P, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant full length protein corresponding to Human Cryptochrome I/CRY1 aa 1-587.

  • General notes

     This product was previously labelled as Cryptochrome I

    This product was changed from ascites to tissue culture supernatant on 4th April 2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

     

Properties

Applications

Our Abpromise guarantee covers the use of ab54649 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use at an assay dependent concentration. Predicted molecular weight: 66 kDa.
ICC/IF Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

Target

  • Function

    Transcriptional repressor 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. CRY1 and CRY2 have redundant functions but also differential and selective contributions at least in defining the pace of the SCN circadian clock and its circadian transcriptional outputs. More potent transcriptional repressor in cerebellum and liver than CRY2, though more effective in lengthening the period of the SCN oscillator. On its side, CRY2 seems to play a critical role in tuning SCN circadian period by opposing the action of CRY1. With CRY2, is dispensable for circadian rhythm generation but necessary for the development of intercellular networks for rhythm synchrony. Capable of translocating circadian clock core proteins such as PER proteins to the nucleus. Interacts with CLOCK-ARNTL/BMAL1 independently of PER proteins and is found at CLOCK-ARNTL/BMAL1-bound sites, suggesting that CRY may act as a molecular gatekeeper to maintain CLOCK-ARNTL/BMAL1 in a poised and repressed state until the proper time for transcriptional activation. Represses the CLOCK-ARNTL/BMAL1 induced transcription of BHLHE40/DEC1. Represses the CLOCK-ARNTL/BMAL1 induced transcription of ATF4, MTA1, KLF10 and NAMPT (By similarity). May repress circadian target genes expression in collaboration with HDAC1 and HDAC2 through histone deacetylation. Mediates the clock-control activation of ATR and modulates ATR-mediated DNA damage checkpoint. In liver, mediates circadian regulation of cAMP signaling and gluconeogenesis by binding to membrane-coupled G proteins and blocking glucagon-mediated increases in intracellular cAMP concentrations and CREB1 phosphorylation. Besides its role in the maintenance of the circadian clock, is also involved in the regulation of other processes. Represses glucocorticoid receptor NR3C1/GR-induced transcriptional activity by binding to glucocorticoid response elements (GREs). Plays a key role in glucose and lipid metabolism modulation, in part, through the transcriptional regulation of genes involved in these pathways, such as LEP or ACSL4.
  • Sequence similarities

    Belongs to the DNA photolyase class-1 family.
    Contains 1 photolyase/cryptochrome alpha/beta domain.
  • Post-translational
    modifications

    Phosphorylation on Ser-247 by MAPK is important for the inhibition of CLOCK-ARNTL/BMAL1-mediated transcriptional activity. Phosphorylation by CSNK1E requires interaction with PER1 or PER2. Phosphorylation at Ser-71 and Ser-280 by AMPK decreases protein stability. Phosphorylation at Ser-568 exhibits a robust circadian rhythm with a peak at CT8, increases protein stability, prevents SCF(FBXL3)-mediated degradation and is antagonized by interaction with PRKDC.
    Ubiquitinated by the SCF(FBXL3) and SCF(FBXL21) complexes, regulating the balance between degradation and stabilization. The SCF(FBXL3) complex is mainly nuclear and mediates ubiquitination and subsequent degradation of CRY1. In contrast, cytoplasmic SCF(FBXL21) complex-mediated ubiquitination leads to stabilize CRY1 and counteract the activity of the SCF(FBXL3) complex. The SCF(FBXL3) and SCF(FBXL21) complexes probably mediate ubiquitination at different Lys residues. Ubiquitination at Lys-11 and Lys-107 are specifically ubiquitinated by the SCF(FBXL21) complex but not by the SCF(FBXL3) complex. Ubiquitination may be inhibited by PER2.
  • Cellular localization

    Cytoplasm. Nucleus. Translocated to the nucleus through interaction with other clock proteins such as PER2 or ARNTL/BMAL1.
  • Information by UniProt
  • Database links

  • Alternative names

    • Cry1 antibody
    • CRY1_HUMAN antibody
    • Cryptochrome 1 (photolyase like) antibody
    • Cryptochrome 1 antibody
    • Cryptochrome-1 antibody
    • PHLL1 antibody
    • Photolyase 1 antibody
    • Photolyase-like antibody
    see all

Images

  • Cryptochrome I/CRY1 antibody (ab54649) at 1ug/lane + HeLa cell lysate at 25ug/lane.

    This image was generated using the ascites version of the product.

  • Cryptochrome I/CRY1 antibody (ab54649) used in immunofluorescence at 10ug/ml on HeLa cells.

    This image was generated using the ascites version of the product.

  • Cryptochrome I/CRY1 antibody (ab54649) used in immunohistochemistry at 5ug/ml on formalin fixed and paraffin embedded human colon adenocarcinoma tissue.

    This image was generated using the ascites version of the product.

  • Overlay histogram showing HeLa cells stained with ab54649 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab54649, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Tween used under the same conditions.

    This image was generated using the ascites version of the product.

References

This product has been referenced in:

See all 5 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Hamster Tissue sections (heart)
Antigen retrieval step
None
Permeabilization
No
Specification
heart
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 21°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Nov 11 2016

Question
Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.
As requested, I have issued a free of charge replacement with ab114543 with the order number x.
To check the status of the order please contact our Customer Service team and reference this number.
Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.
I wish you the best of luck with your research.

Read More

Answer

Thank you for contacting us.
I am sorry to hear you have been experiencing problems with our Cryptochrome antibodies. The quality of our products is important to us and I would like to reassure you that we investigate all customer complaints.
We have received no complaints about these products and I am sorry that you doubt the specificity. As both Cryptochrome proteins are post-translationally modified for example by phosphorylation, this could influence the electrophoretic mobility of the proteins in the gel. The observed band size may also dependent on other factors such as the buffer system used. For an example see the datasheet of ab93802. Considering this, the bands you observed seem specific to me.
In order to optimise the results and reduce the background staining I would recommend the following:
1) As the cryptochromes translocate to the nucleus it might be useful to analyse the cell fractions cytomplasm and nucleus separately. Protocol details can be found on our website (https://www.abcam.com/index.html?pageconfig=popular_protocols).
2) When testing our antibodies, our lab uses 5% BSA as a blocking reagent, so I recommend switching to this instead of milk, if you have not used it already. For unknown reasons some antibodies give stronger, more specific signals on blots blocked with BSA instead of milk, so doing this may improve the results you are seeing, and reduce the non-specific bands.
3) The amount of protein loaded per lane and the concentration of the primary antibody might be optimised, too, as the bands are fairly strong.
As the results appear to be specific and thus the antibodies seem to work, we would usually not send a free of charge control sample. However, in this particular case I am authorised to make an exception and could send you our Cryptochrome I protein ab114543:
https://www.abcam.com/index.html?datasheet=114543 https://www.abcam.com/index.html?datasheet=114543.
We unfortunately do not have a suitable Cryptochrome II protein available, but I hope using ab114543 will help to validate the results.
If you would like to accept this offer, could you please confirm the order details for me so that I could arrange for the shipment?
order number 1018633 (PO135) from 30 Jan 2012
lot number 1230051/GR47664-2 (ab54649) and 1235524 GR71422-2 (ab119053)
I look forward to your reply.

Read More
Application
Western blot
Sample
Mouse Tissue lysate - whole (brain)
Gel Running Conditions
Reduced Denaturing (10% gel)
Loading amount
25 µg
Specification
brain
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Oct 31 2011

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (pooled primary human keratinocytes)
Permeabilization
Yes - 0.1% Triton-X-100
Specification
pooled primary human keratinocytes
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Paraformaldehyde

Dr. Joerg Liebmann

Verified customer

Submitted May 18 2010

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up