Invitrogen

CLOCK Polyclonal Antibody

Advanced Verification

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

3 Published Figures

10 References

View all (28) CLOCK antibodies

Datasheet

Questions & Answers

Datasheet

Questions & Answers

Product Image Gallery

CLOCK Antibody in Immunofluorescence (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 in Relative expression — 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 — Published figure using CLOCK polyclonal antibody (Product # PA1-520)

Product Details

Applications

Tested Dilution

Publications

Immunocytochemistry (ICC)

1:10-1:200

View 1 publication 1 publication

Immunofluorescence (IF)

1:10-1:200

View 1 publication 1 publication

Immunohistochemistry (Frozen) (IHC (F))

1:100-1:1000

Immunohistochemistry (Paraffin) (IHC (P))

1:100-1:1000

Western Blot (WB)

1:200-1:2000

View 3 publications 3 publications

Immunohistochemistry - Free Floating (IHC (Free))

View 1 publication 1 publication

Gel Shift (GS)

View 1 publication 1 publication

Immunohistochemistry (IHC)

View 3 publications 3 publications

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.

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

Molecular Function: basic helix-loop-helix transcription factor transcription factor