Anti-KAT13D / CLOCK抗体- ChIP Grade (ab3517)

概述

  • 产品名称Anti-KAT13D / CLOCK抗体- ChIP Grade
    参阅全部 KAT13D / CLOCK 一抗
  • 描述
    兔多克隆抗体to KAT13D / CLOCK - ChIP Grade
  • 经测试应用适用于: IHC-P, IHC-Fr, WB, ChIP, ICC/IFmore details
  • 种属反应性
    与反应: Mouse, Rat, Hamster, Human
  • 免疫原

    Synthetic peptide corresponding to Mouse KAT13D/ CLOCK aa 11-30.
    Sequence:

    SSIVDRDDSSIFDGLVEEDD


    (Peptide available as ab5853)

应用

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.

应用 Ab评论 说明
IHC-P 1/100 - 1/1000.

Immunohistochemical staining of CLOCK in hamster brain results in the staining of the superchiasmatic nucleus.

EMSA Use at an assay dependent concentration. PubMed: 22210883
IHC-Fr 1/100 - 1/1000.
WB 1/200. Detects a band of approximately 100 kDa (predicted molecular weight: 95 kDa).Can be blocked with PAR4 peptide (ab5853). By Western blot, this antibody detects an ~100 kDa protein representing CLOCK from rat brain homogenate.
ChIP Use at an assay dependent concentration. PubMed: 20956306
ICC/IF Use at an assay dependent concentration. PubMed: 22028779

靶标

  • 功能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.
  • 组织特异性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).
  • 序列相似性Contains 1 basic helix-loop-helix (bHLH) domain.
    Contains 1 PAC (PAS-associated C-terminal) domain.
    Contains 2 PAS (PER-ARNT-SIM) domains.
  • 翻译后修饰Phosphorylation is dependent on CLOCK-ARNTL heterodimer formation.
  • 细胞定位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
  • 数据库链接
  • 别名
    • bHLHe8 antibody
    • Circadian locomoter output cycles kaput protein antibody
    • Circadian locomoter output cycles protein kaput antibody
    • Circadian Locomotor Output Cycles Kaput antibody
    • Circadium Locomotor Output Cycles Kaput antibody
    • Class E basic helix-loop-helix protein 8 antibody
    • CLOCK antibody
    • Clock circadian regulator antibody
    • Clock homolog antibody
    • Clock protein antibody
    • CLOCK_HUMAN antibody
    • hCLOCK antibody
    • KIAA0334 antibody
    see all

Anti-KAT13D / CLOCK antibody - ChIP Grade 图像

  • 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.

  • 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.

  • 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.

  • 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.

  • ab3517 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.

    See Abreview

  • All lanes : Anti-KAT13D / CLOCK antibody - ChIP Grade (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?)
  • All lanes : Anti-KAT13D / CLOCK antibody - ChIP Grade (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
    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.

Anti-KAT13D / CLOCK antibody - ChIP Grade (ab3517)参考文献

This product has been referenced in:
  • Meier D  et al. Twist1 Is a TNF-Inducible Inhibitor of Clock Mediated Activation of Period Genes. PLoS One 10:e0137229 (2015). ChIP . Read more (PubMed: 26361389) »
  • Mori K  et al. Epigenetic suppression of mouse Per2 expression in the suprachiasmatic nucleus by the inhalational anesthetic, sevoflurane. PLoS One 9:e87319 (2014). ChIP ; Mouse . Read more (PubMed: 24498074) »

See all 11 Publications for this product

Product Wall

Application Western blot
Sample Human Cell lysate - whole cell (human fibroblast cells)
Gel Running Conditions Reduced Denaturing (7.5%)
Loading amount 20 µg
Specification human fibroblast cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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Abcam user community

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提交于 Oct 28 2015

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (hepatocytes)
Permeabilization Yes - 0.1% Triton X-100 in TBS for 5-10 minutes
Specification hepatocytes
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 23°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

提交于 Sep 11 2015

Thank you for contacting us.
I am sorry to hear that the antibody has not been working as expected.

As we discussed over the phone, trying a dilution of 1/500 or 1/1000 might help to improve your results.

Please let me know if...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (brain)
Loading amount 25 µg
Specification brain
Gel Running Conditions Reduced Denaturing (10% gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Jul 19 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Skeletal muscle tissue)
Specification Skeletal muscle tissue
Fixative Paraformaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric buffer, pH 6
Permeabilization No
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
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提交于 Mar 18 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (Lung homogenate)
Loading amount 100 µg
Specification Lung homogenate
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5%
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提交于 Apr 06 2007

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"