Invitrogen

CLOCK Polyclonal Antibody

Advanced Verification

This Antibody was verified by Relative expression to ensure that the antibody binds to the antigen stated.

6 Published Figures

10 References

View all (30) CLOCK antibodies

Datasheet

Protocols

Questions & Answers

Tech Support

Datasheet

Protocols

Questions & Answers

Tech Support

Product Image Gallery

CLOCK Antibody in Immunocytochemistry (ICC/IF) — 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 fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

CLOCK Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of CLOCK showing positive staining in the nucleus and cytoplasm of paraffin-treated Human skeletal muscle (right) compared with a negative control in the absence of primary antibody (left). To expose target proteins, antigen retrieval method was performed using 10mM sodium citrate (pH 6.0) microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a CLOCK polyclonal antibody (Product # PA1-520) diluted by 3% BSA-PBS at a dilution of 1:200 overnight at 4°C in a humidified chamber. Tissues were washed extensively PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

CLOCK Antibody in Western Blot (WB) — Western blot was performed using Anti-CLOCK Polyclonal Antibody (Product # PA1-520) and a ~95 kDa band corresponding to CLOCK was observed in Mouse Skeletal Muscle and not in Mouse Liver which is reported to be negative. An uncharacterized band (*) at ~260 kDa was observed in Mouse Skeletal Muscle. Tissue extracts (30ug) of Mouse Skeletal Muscle (Lane 1), Mouse Liver (Lane 2) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:2000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

CLOCK Antibody — Antibody specificity was demonstrated by detection of differential basal expression of the target across tissues owing to their inherent genetic constitution. SATB1 was observed in Mouse Skeletal Muscle but not in Mouse Liver using CLOCK Polyclonal Antibody (Product # PA1-520) in western blot. {RE}

CLOCK Antibody in Immunohistochemistry (IHC) — Published figure using CLOCK polyclonal antibody (Product # PA1-520) in Immunohistochemistry

Product Details

PA1-520

Applications	Tested Dilution	Publications
Western Blot (WB)	1:200-1:2,000	View 3 publications 3 publications
Immunohistochemistry (IHC)	-	View 4 publications 4 publications
Immunohistochemistry (Paraffin) (IHC (P))	1:100-1:1,000	-
Immunohistochemistry (Frozen) (IHC (F))	1:100-1:1,000	-
Immunohistochemistry - Free Floating (IHC (Free))	-	View 1 publication 1 publication
Immunocytochemistry (ICC/IF)	1:10-1:200	View 1 publication 1 publication
Gel Shift (GS)	-	View 1 publication 1 publication

Product Specifications
Species Reactivity	Hamster, Human, Mouse, Rat
Published species	Human, Mouse, Rat
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
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
Conjugate	Unconjugated
Form	Liquid
Concentration	1 mg/mL
Purification	Antigen affinity chromatography
Storage buffer	PBS with 1mg/mL BSA
Contains	0.05% sodium azide
Storage conditions	-20° C, Avoid Freeze/Thaw Cycles
RRID	AB_2082586

Product Specific Information

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.

Target Information

This gene encodes a protein that belongs to the basic helix-loop-helix (bHLH) family of transcription factors. Polymorphisms within the encoded protein have been associated with circadian rhythm sleep disorders. A similar protein in mice is a circadian regulator that acts as a transcription factor and forms a heterodimer with aryl hydrocarbon receptor nuclear translocator-like to activate transcription of mouse period 1.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

Melatonin modulates daytime-dependent synaptic plasticity and learning efficiency.

Journal of pineal research

Jilg A,Bechstein P,Saade A,Dick M,Li TX,Tosini G,Rami A,Zemmar A,Stehle JH

PA1-520 was used in Immunohistochemistry to investigate the role of the neurohormone melatonin as a circadian time cue for hippocampal signaling and memory formation.

Mon Apr 01 00:00:00 EDT 2019

Comparative cell cycle transcriptomics reveals synchronization of developmental transcription factor networks in cancer cells.

PloS one

Boström J,Sramkova Z,Salašová A,Johard H,Mahdessian D,Fedr R,Marks C,Medalová J,Souček K,Lundberg E,Linnarsson S,Bryja V,Sekyrova P,Altun M,Andäng M

PA1-520 was used in Immunocytochemistry-immunoflourescence to characterise the transcriptomes of unsynchronised HeLa and U2OS cancer cells, sorted for cell cycle phase by Fucci reporter expression.

Thu Jan 04 00:00:00 EST 2018

Time-caloric restriction inhibits the neoplastic transformation of cirrhotic liver in rats treated with diethylnitrosamine.

Carcinogenesis

Molina-Aguilar C,Guerrero-Carrillo MJ,Espinosa-Aguirre JJ,Olguin-Reyes S,Castro-Belio T,Vázquez-Martínez O,Rivera-Zavala JB,Díaz-Muñoz M

PA1-520 was used in Western Blotting to conclude that time-caloric restriction prevents fibrosis from progressing into cirrhosis, thus avoiding chronic inflammation and regenerative processes.

Tue Aug 01 00:00:00 EDT 2017

Circadian networks in human embryonic stem cell-derived cardiomyocytes.

EMBO reports

Dierickx P,Vermunt MW,Muraro MJ,Creyghton MP,Doevendans PA,van Oudenaarden A,Geijsen N,Van Laake LW

Published figure using CLOCK polyclonal antibody (Product # PA1-520) in Western Blot

Sat Jul 01 00:00:00 EDT 2017

Circadian networks in human embryonic stem cell-derived cardiomyocytes.

EMBO reports

Dierickx P,Vermunt MW,Muraro MJ,Creyghton MP,Doevendans PA,van Oudenaarden A,Geijsen N,Van Laake LW

Published figure using CLOCK polyclonal antibody (Product # PA1-520) in Western Blot

Sat Jul 01 00:00:00 EDT 2017

Diurnal oscillation of amygdala clock gene expression and loss of synchrony in a mouse model of depression.

The international journal of neuropsychopharmacology

Savalli G,Diao W,Schulz S,Todtova K,Pollak DD

PA1-520 was used in immunohistochemistry - free floating to discuss the contribution of the circadian system to depression.

Thu Dec 11 00:00:00 EST 2014

p53 regulates Period2 expression and the circadian clock.

Nature communications

Miki T,Matsumoto T,Zhao Z,Lee CC

PA1-520 was used in EMSA to assess the role of p53 in regulating Period2 expression and the circadian clock

Mon Apr 21 00:00:00 EDT 2014

Circadian rhythms regulate amelogenesis.

Bone

Zheng L,Seon YJ,Mourão MA,Schnell S,Kim D,Harada H,Papagerakis S,Papagerakis P

PA1-520 was used in western blot to study the circadian regulation of amelogenesis

Mon Jul 01 00:00:00 EDT 2013

Expression of clock proteins in developing tooth.

Gene expression patterns : GEP

Zheng L,Papagerakis S,Schnell SD,Hoogerwerf WA,Papagerakis P

PA1-520 was used in immunohistochemistry to study the role of clock genes in developing tooth

Thu Jul 21 00:00:00 EDT 2011

The E2F-regulated gene Chk1 is highly expressed in triple-negative estrogen receptor /progesterone receptor /HER-2 breast carcinomas.

Cancer research

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

PA1-520 was used in immunohistochemistry to study the expression of Chk1 in triple-negative estrogen receptor/progesterone receptor/HER-2 breast carcinomas

Sun Jul 15 00:00:00 EDT 2007

Non-cyclic and developmental stage-specific expression of circadian clock proteins during murine spermatogenesis.

Biology of reproduction

Alvarez JD,Chen D,Storer E,Sehgal A

PA1-520 was used in immunohistochemistry to investigate the expression of circadian clock proteins during murine spermatogenesis

Tue Jul 01 00:00:00 EDT 2003

Bioinformatics

Protein Aliases: bHLHe8; circadian locomoter output cycles kaput; circadian locomoter output cycles kaput protein; Circadian locomoter output cycles protein kaput; Class E basic helix-loop-helix protein 8; clock circadian regulator; clock homolog; hCLOCK; mCLOCK; rCLOCK

Gene Aliases: 5330400M04Rik; BHLHE8; CLOCK; I79_020252; KAT13D; KIAA0334

UniProt ID: (Human) O15516, (Mouse) O08785, (Rat) Q9WVS9

Entrez Gene ID: (Human) 9575, (Chinese hamster) 100770092, (Mouse) 12753, (Rat) 60447

Function(s)