Anti-KAT13D / CLOCK antibody (ab3517)
Key features and details
- Rabbit polyclonal to KAT13D / CLOCK
- Suitable for: ICC, IHC-P, WB
- Reacts with: Mouse, Rat, Hamster
- Isotype: IgG
Overview
-
Product name
Anti-KAT13D / CLOCK antibody
See all KAT13D / CLOCK primary antibodies -
Description
Rabbit polyclonal to KAT13D / CLOCK -
Host species
Rabbit -
Tested applications
Suitable for: ICC, IHC-P, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Hamster -
Immunogen
Synthetic peptide:
SSIVDRDDSSIFDGLVEEDD
, corresponding to amino acids 11-30 of Mouse KAT13D/ CLOCK(Peptide available as ab5854.)
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 99% PBS -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
-
ChIP Related Products
-
Compatible Secondaries
-
Isotype control
-
Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab3517 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 |
|---|---|---|
| ICC | Use at an assay dependent concentration. | |
| IHC-P | 1/100 - 1/1000. Immunohistochemical staining of CLOCK in hamster brain results in the staining of the superchiasmatic nucleus. |
|
| WB | 1/200. Detects a band of approximately 100 kDa (predicted molecular weight: 95 kDa). By Western blot, this antibody detects an ~100 kDa protein representing CLOCK from rat brain homogenate. |
Target
-
Function
ARNTL/2-CLOCK heterodimers activate E-box element (3'-CACGTG-5') transcription of a number of proteins of the circadian clock. Activates transcription of PER1 and PER2. This transcription is inhibited in a feedback loop by PER and CRY proteins. Has intrinsic histone acetyltransferase activity and this enzymatic function contributes to chromatin-remodeling events implicated in circadian control of gene expression (By similarity). Acetylates primarily histones H3 and H4 (By similarity). Acetylates also a non-histone substrate: ARNTL. -
Tissue specificity
Expressed in all tissues examined including spleen, thymus, prostate, testis, ovary, small intestine, colon, leukocytes, heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas. Highest levels in testis and skeletal muscle. Low levels in thymus, lung and liver. Expressed in all brain regions with highest levels in cerebellum. Highly expressed in the suprachiasmatic nucleus (SCN). -
Sequence similarities
Contains 1 basic helix-loop-helix (bHLH) domain.
Contains 1 PAC (PAS-associated C-terminal) domain.
Contains 2 PAS (PER-ARNT-SIM) domains. -
Post-translational
modificationsPhosphorylation is dependent on CLOCK-ARNTL heterodimer formation. -
Cellular localization
Cytoplasm. Nucleus. Shuffling between the cytoplasm and the nucleus is under circadian regulation and is ARNTL-dependent. Phosphorylated form located in the nucleus. - Information by UniProt
-
Database links
- Entrez Gene: 12753 Mouse
- Entrez Gene: 60447 Rat
- SwissProt: O08785 Mouse
- SwissProt: Q9WVS9 Rat
- Unigene: 3552 Mouse
- Unigene: 392894 Mouse
- Unigene: 205839 Rat
-
Alternative names
- bHLHe8 antibody
- Circadian locomoter output cycles kaput protein antibody
- Circadian locomoter output cycles protein kaput antibody
see all
Images
-
Immunocytochemistry - Anti-KAT13D / CLOCK antibody (ab3517)Immunocytochemistry/immunofluorescence analysis of U251 cells labeling KAT13D/CLOCK (green) with ab3517 at 1/100. Cells were fixed with formalin and permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blcoked with £% BSA in PBS for 30 minutes at room temperature. Cells were incubated with the primary antibody overnight at 4°C. A DyLight-conjugated secondary antibody was used. F-actin (red) was stained with phalloidin and nuclei (blue) were stained with Hoechst or DAPI. 60X magnification. Left - negative control.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KAT13D / CLOCK antibody (ab3517)ab3517 labelling KAT13D in the nucleus and cytoplasm of Human skeletal muscle tissue (right) compared with a negative control (left) by Immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections). 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. Thissue sections were incubated with the primary antibody (1:200 in 3% BSA-PBS) overnight at 4°C. A HRP-conjugated anti-rabbit IgG was used as the 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.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KAT13D / CLOCK antibody (ab3517)ab3517 labelling KAT13D in the nucleus and cytoplasm of Mouse colon tissue (right) compared with a negative control (left) by Immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections). 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. Thissue sections were incubated with the primary antibody (1:200 in 3% BSA-PBS) overnight at 4°C. A HRP-conjugated anti-rabbit IgG was used as the 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.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KAT13D / CLOCK antibody (ab3517)ab3517 labelling KAT13D in the nucleus and cytoplasm of Human colon tissue (right) compared with a negative control (left) by Immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections). 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. Thissue sections were incubated with the primary antibody (1:200 in 3% BSA-PBS) overnight at 4°C. A HRP-conjugated anti-rabbit IgG was used as the 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.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KAT13D / CLOCK antibody (ab3517)This image is courtesy of an anonymous Abreviewab3517 staining KAT13D/CLOCK in Mouse skeletal muscle tissue sections by IHC-P (Paraformaldehyde-fixed, paraffin-embedded tissue sections). Tissue was fixed with paraformaldehyde and blocked with 10% serum for 1 hour at 20°C; antigen retrieval was by heat mediation in citrate buffer pH6. Samples were incubated with primary antibody (1/400 in PBS) for 12 hours at 4°C. Undiluted ab64256 was used as the secondary antibody. -
Western blot - Anti-KAT13D / CLOCK antibody (ab3517)All lanes : Anti-KAT13D / CLOCK antibody (ab3517) at 1/4000 dilution
Lane 1 : HeLa cell lysate
Lane 2 : NIH-3T3 cell lysate
Lysates/proteins at 25 µg per lane.
Predicted band size: 95 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted? -
Western blot - Anti-KAT13D / CLOCK antibody (ab3517)All lanes : Anti-KAT13D / CLOCK antibody (ab3517) at 1 µg/ml
Lane 1 : Skeletal Muscle (Mouse) Tissue Lysate
Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 95 kDa
Observed band size: 95 kDa
Additional bands at: 105 kDa (possible post-translational modification), 38 kDa, 45 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 30 seconds
KAT13D / CLOCK contains a number of potential phosphorylation sites (SwissProt) which may explain the banding pattern observed at a higher molecular weight than predicted.
Protocols
References (26)
ab3517 has been referenced in 26 publications.
- Yao Y et al. Noninvasive 40-Hz Light Flicker Rescues Circadian Behavior and Abnormal Lipid Metabolism Induced by Acute Ethanol Exposure via Improving SIRT1 and the Circadian Clock in the Liver-Brain Axis. Front Pharmacol 11:355 (2020). PubMed: 32269528
- Chen L et al. Pinealectomy or light exposure exacerbates biliary damage and liver fibrosis in cholestatic rats through decreased melatonin synthesis. Biochim Biophys Acta Mol Basis Dis 1865:1525-1539 (2019). PubMed: 30890428
- Fader KA et al. 2,3,7,8-Tetrachlorodibenzo-p-dioxin abolishes circadian regulation of hepatic metabolic activity in mice. Sci Rep 9:6514 (2019). PubMed: 31015483
- Castelo-Szekely V et al. Charting DENR-dependent translation reinitiation uncovers predictive uORF features and links to circadian timekeeping via Clock. Nucleic Acids Res 47:5193-5209 (2019). PubMed: 30982898
- Minegishi S et al. Circadian clock disruption by selective removal of endogenous carbon monoxide. Sci Rep 8:11996 (2018). PubMed: 30097595
