Anti-Casein Kinase 1 delta antibody [AF12G4] (ab85320)

Overview

  • Product nameAnti-Casein Kinase 1 delta antibody [AF12G4]
    See all Casein Kinase 1 delta primary antibodies
  • Description
    Mouse monoclonal [AF12G4] to Casein Kinase 1 delta
  • Tested applicationsSuitable for: IHC-P, Flow Cyt, WB, IP, ELISAmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    His tagged recombinant Human Casein Kinase 1 delta protein, purified from E.coli.

  • Positive control
    • HeLa, K562, SK-N-MC, 293T and L929 cell lysates. This antibody also gave a positive signal in Human Alzheimer brain (hippocampus) tissue section.

Properties

Applications

Our Abpromise guarantee covers the use of ab85320 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 Use a concentration of 0.2 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Flow Cyt Use 1µg for 106 cells. ab170192-Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.
WB 1/5000. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).
IP Use at an assay dependent concentration.
ELISA 1/5000.

Target

  • FunctionCasein kinases are operationally defined by their preferential utilization of acidic proteins such as caseins as substrates. It can phosphorylate a large number of proteins. Participates in Wnt signaling. Central component of the circadian clock. May act as a negative regulator of circadian rhythmicity by phosphorylating PER1 and PER2. Retains PER1 in the cytoplasm.
  • Tissue specificityExpressed in all tissues examined, including brain, heart, lung, liver, pancreas, kidney, placenta and skeletal muscle. In blood, highly expressed in hemopoietic cells and mature granulocytes. Also found in monocytes and lymphocytes.
  • Involvement in diseaseDefects in CSNK1D are a cause of familial advanced sleep-phase syndrome (FASPS) [MIM:604348]. FASPS is characterized by very early sleep onset and offset. Individuals are 'morning larks' with a 4 hours advance of the sleep, temperature and melatonin rhythms.
  • Sequence similaritiesBelongs to the protein kinase superfamily. CK1 Ser/Thr protein kinase family. Casein kinase I subfamily.
    Contains 1 protein kinase domain.
  • Post-translational
    modifications
    Autophosphorylated on serine and threonine residues.
  • Cellular localizationCytoplasm. Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • Casein kinase I delta isoform antibody
    • Casein kinase I isoform delta antibody
    • CKI delta antibody
    • CKI-delta antibody
    • CKId antibody
    • CKIdelta antibody
    • CSNK 1D antibody
    • CSNK1D antibody
    • HCKID antibody
    • KC1D_HUMAN antibody
    • Protein kinase CK1 delta antibody
    see all

Anti-Casein Kinase 1 delta antibody [AF12G4] images

  • All lanes : Anti-Casein Kinase 1 delta antibody [AF12G4] (ab85320) at 1/5000 dilution

    Lane 1 : HeLa cell lysate
    Lane 2 : K562 cell lysate
    Lane 3 : SK-N-MC cell lysate
    Lane 4 : 293T cell lysate
    Lane 5 : L929 cell lysate


    Predicted band size : 47 kDa
    Observed band size : 47 kDa
  • Overlay histogram showing HeLa cells stained with ab85320 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab85320, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • IHC image of ab85320 staining in human Alzheimer brain (hippocampus) formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab85320, 0.2µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
  • Casein Kinase 1 delta was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Mouse monoclonal to Casein Kinase 1 delta and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab85320.
    Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
    Band: 47kDa: Casein Kinase 1 delta; non specific - 40kDa: We are unsure as to the identity of this extra band.

References for Anti-Casein Kinase 1 delta antibody [AF12G4] (ab85320)

ab85320 has not yet been referenced specifically in any publications.

Product Wall

Application Western blot
Sample Mouse Purified protein (liver)
Gel Running Conditions Non-reduced Non-Denaturing (Native) (4-16% Bis-Tris)
Loading amount 0.001 µg
Specification liver
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 1%
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Mariska Twaalfhoven

Verified customer

Submitted Dec 18 2015

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