Immunofluorescent analysis of CLOCK (green) showing staining in the nucleus and cytoplasm of U251 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a CLOCK polyclonal antibody (Product # PA1-520) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 °C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a flourescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.
|Tested species reactivity||Hamster, Human, Mouse, Rat|
|Published species reactivity||Mouse, Human, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic Peptide: S(11) S I V D R D D S S I F D G L V E E D D(30) C|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunohistochemistry (Frozen) (IHC (F))||1:100-1:1000|
|Immunohistochemistry (Paraffin) (IHC (P))||1:100-1:1000|
|Western Blot (WB)||1:200|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PA1-520 detects CLOCK from human, mouse, rat and hamster tissues.
PA1-520 has been successfully used in Western blot, ICC/IF and immunohistochemistry procedures. By Western blot, this antibody detects an ~100 kDa protein representing CLOCK from rat brain homogenate. Immunohistochemical staining of CLOCK in hamster brain with PA1-520 results in the staining of the superchiasmatic nucleus.
PA1-520 immunizing peptide corresponds to amino acid residues 11-30 of mouse CLOCK. This sequence is completely conserved between mouse and human CLOCK. PA1-520 immunizing peptide (Cat. # PEP-072) is available for use in neutralization and control experiments.
Circadian rhythmicity is a basic property of phylogenetically diverse organisms which range from animals and plants, to fungi. Regulation of endogenous biological clocks is regulated at the genetic level by a protein-mediated, autoregulatory feed-back loop. In mammals, several genes that encode members of the basic helix-loop helix (bHLH) PAS (PER-ARNT-SIM) transcription factor family have been shown to play a significant role in regulating circadian oscillations. Transactivation of CLOCK-induced genes is mediated via an E box enhancer (CACGTG) found upstream of target genes. CLOCK-ARNT3 heterodimers bind to E box regulatory elements and stimulate gene transcription. CLOCK has been shown to transactivate the mammalian homolog of Drosophila per. PER, in concert with the product of the mammalian timeless gene (TIM), negatively regulates its own transcription by blocking the activity of the CLOCK-BMAL1 transactivation complex.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Diurnal oscillation of amygdala clock gene expression and loss of synchrony in a mouse model of depression.
PA1-520 was used in immunohistochemistry - free floating to discuss the contribution of the circadian system to depression.
|Savalli G,Diao W,Schulz S,Todtova K,Pollak DD||The international journal of neuropsychopharmacology (18:null)||2014|
|Not Applicable||Not Cited||
p53 regulates Period2 expression and the circadian clock.
PA1-520 was used in EMSA to assess the role of p53 in regulating Period2 expression and the circadian clock
|Miki T,Matsumoto T,Zhao Z,Lee CC||Nature communications (4:null)||2013|
Circadian rhythms regulate amelogenesis.
PA1-520 was used in western blot to study the circadian regulation of amelogenesis
|Zheng L,Seon YJ,Mourão MA,Schnell S,Kim D,Harada H,Papagerakis S,Papagerakis P||Bone (55:158)||2013|
Expression of clock proteins in developing tooth.
PA1-520 was used in immunohistochemistry to study the role of clock genes in developing tooth
|Zheng L,Papagerakis S,Schnell SD,Hoogerwerf WA,Papagerakis P||Gene expression patterns : GEP (11:202)||2011|
The E2F-regulated gene Chk1 is highly expressed in triple-negative estrogen receptor /progesterone receptor /HER-2 breast carcinomas.
PA1-520 was used in immunohistochemistry to study the expression of Chk1 in triple-negative estrogen receptor/progesterone receptor/HER-2 breast carcinomas
|Verlinden L,Vanden Bempt I,Eelen G,Drijkoningen M,Verlinden I,Marchal K,De Wolf-Peeters C,Christiaens MR,Michiels L,Bouillon R,Verstuyf A||Cancer research (67:6574)||2007|
Non-cyclic and developmental stage-specific expression of circadian clock proteins during murine spermatogenesis.
PA1-520 was used in immunohistochemistry to investigate the expression of circadian clock proteins during murine spermatogenesis
|Alvarez JD,Chen D,Storer E,Sehgal A||Biology of reproduction (69:81)||2003|