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    kat13d--clock-antibody-ab3517.pdf

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Anti-KAT13D / CLOCK antibody (ab3517)

  • Datasheet
  • SDS
Reviews (5)Q&A (1)References (41)

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Western blot - Anti-KAT13D / CLOCK antibody (ab3517)
  • Immunocytochemistry/ Immunofluorescence - Anti-KAT13D / CLOCK antibody (ab3517)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KAT13D / CLOCK antibody (ab3517)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KAT13D / CLOCK antibody (ab3517)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KAT13D / CLOCK antibody (ab3517)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KAT13D / CLOCK antibody (ab3517)
  • Western blot - Anti-KAT13D / CLOCK antibody (ab3517)

Key features and details

  • Rabbit polyclonal to KAT13D / CLOCK
  • Suitable for: IHC-P, ICC/IF, IHC-Fr, WB, ChIP
  • Reacts with: Mouse, Human
  • Isotype: IgG

You may also be interested in

Protein
Product image
Recombinant Human KAT13D / CLOCK protein (ab153001)
Primary
Product image
Anti-PER2 antibody [EPR11381(2)] (ab179813)
Secondary
Product image
Goat Anti-Rabbit IgG H&L (HRP) (ab205718)

View more associated products

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: IHC-P, ICC/IF, IHC-Fr, WB, ChIPmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide corresponding to Mouse KAT13D/ CLOCK aa 1-100.

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • Positive control

    • WB: human HeLa, mouse NIH-3T3 and skeletal muscle tissue; IHC: human skeletal muscle, colon tissue, mouse colon tissue; ICC: human U251 cells
  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

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

    • Neuroscience
    • Neurology process
    • Circadian Rhythm
    • Signal Transduction
    • Metabolism
    • Energy Metabolism
    • Neuroscience
    • Neurology process
    • Metabolism
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Transcription Factors
    • Epigenetics and Nuclear Signaling
    • Chromatin Modifying Enzymes
    • Acetylation
    • HAT
    • Cardiovascular
    • Atherosclerosis
    • Lipoprotein metabolism
    • Cardiovascular
    • Blood
    • Blood Pressure regulation
    • Cardiovascular
    • Heart
    • Cardiac metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Lipid and lipoprotein metabolism
    • Lipoprotein metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Energy transfer pathways
    • Energy Metabolism
    • Metabolism
    • Types of disease
    • Cancer
    • Metabolism
    • Types of disease
    • Heart disease

Associated products

  • ChIP Related Products

    • ChIP Kit (ab500)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human KAT13D / CLOCK protein (ab153001)

Applications

The Abpromise guarantee

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
IHC-P (1)
1/100 - 1/1000.

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

ICC/IF (1)
1/10 - 1/200.
EMSA
Use at an assay dependent concentration.
IHC-Fr
Use at an assay dependent concentration.
Gel Shift Assay
Use at an assay dependent concentration.
WB (3)
1/200 - 1/2000. Detects a band of approximately 100 kDa (predicted molecular weight: 95 kDa).
ChIP
Use at an assay dependent concentration. PubMed: 20956306
Notes
IHC-P
1/100 - 1/1000.

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

ICC/IF
1/10 - 1/200.
EMSA
Use at an assay dependent concentration.
IHC-Fr
Use at an assay dependent concentration.
Gel Shift Assay
Use at an assay dependent concentration.
WB
1/200 - 1/2000. Detects a band of approximately 100 kDa (predicted molecular weight: 95 kDa).
ChIP
Use at an assay dependent concentration. PubMed: 20956306

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
    modifications

    Phosphorylation 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.
  • Target information above from: UniProt accession O15516 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 9575 Human
    • Entrez Gene: 12753 Mouse
    • Omim: 601851 Human
    • SwissProt: O15516 Human
    • SwissProt: O08785 Mouse
    • Unigene: 436975 Human
    • Unigene: 3552 Mouse
    • Unigene: 392894 Mouse
    • Alternative names

      • 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

    Images

    • Western blot - Anti-KAT13D / CLOCK antibody (ab3517)
      Western blot - Anti-KAT13D / CLOCK antibody (ab3517)
      All lanes : Anti-KAT13D / CLOCK antibody (ab3517) at 1/2000 dilution

      Lane 1 : Mouse skeletal muscle tissue
      Lane 2 : Mouse liver tissue

      Lysates/proteins at 30 µg per lane.

      Secondary
      Lane 1 : Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP at 1/4000 dilution
      Lane 2 : oat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP at 1/4000 dilution

      Developed using the ECL technique.

      Predicted band size: 95 kDa



      Electrophoresis performed on a 4-12% BisTris gel and proteins transferred onto a nitrocellulose membrane.

    • Immunocytochemistry/ Immunofluorescence - Anti-KAT13D / CLOCK antibody (ab3517)
      Immunocytochemistry/ Immunofluorescence - 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)
      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)
      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)
      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)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KAT13D / CLOCK antibody (ab3517)This image is courtesy of an anonymous Abreview
      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

    • Western blot - Anti-KAT13D / CLOCK antibody (ab3517)
      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?

    Protocols

    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (41)

    Publishing research using ab3517? Please let us know so that we can cite the reference in this datasheet.

    ab3517 has been referenced in 41 publications.

    • Park MW  et al. NOX4 promotes ferroptosis of astrocytes by oxidative stress-induced lipid peroxidation via the impairment of mitochondrial metabolism in Alzheimer's diseases. Redox Biol 41:101947 (2021). PubMed: 33774476
    • Ogino T  et al. Post-transcriptional repression of circadian component CLOCK regulates cancer-stemness in murine breast cancer cells. Elife 10:N/A (2021). PubMed: 33890571
    • Yoshida Y  et al. Alteration of circadian machinery in monocytes underlies chronic kidney disease-associated cardiac inflammation and fibrosis. Nat Commun 12:2783 (2021). PubMed: 33986294
    • 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
    • Yao Y  et al. Non-invasive 40-Hz Light Flicker Ameliorates Alzheimer's-Associated Rhythm Disorder via Regulating Central Circadian Clock in Mice. Front Physiol 11:294 (2020). PubMed: 32390857
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-6 of 6 Abreviews or Q&A

    Western blot abreview for Anti-KAT13D / CLOCK antibody - ChIP Grade

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 28 2015

    Immunocytochemistry/ Immunofluorescence abreview for Anti-KAT13D / CLOCK antibody - ChIP Grade

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 11 2015

    Western blot abreview for Anti-KAT13D / CLOCK antibody - ChIP Grade

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 19 2011

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-KAT13D / CLOCK antibody - ChIP Grade

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Mar 18 2010

    Western blot abreview for Anti-KAT13D / CLOCK antibody - ChIP Grade

    Good
    Abreviews
    Abreviews
    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%
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 06 2007

    Question

    mouse heart samples
    ChIP with demo, clock and isotype crtl
    detection by RT-PCR
    pos. crtl: no signal
    clock: same signal like neg. crtl
    demo: good signal
    Ab: 1/50 , 1/5000 (10 ul Ab, 500 ul chromatin)

    suggestion: try 1/500 or 1/1000 as well, check publication which mentioned ChIP

    Read More

    Abcam community

    Verified customer

    Asked on Aug 28 2012

    Answer

    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 this suggestion is of help.

    I look forward to hear back from you.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Abcam Scientific Support

    Answered on Aug 28 2012

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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