Key features and details
- Mouse monoclonal [AF12G4] to Casein Kinase 1 delta/CSNK1D
- Suitable for: IHC-P, Flow Cyt, WB, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
- Isotype: IgG2b
Product nameAnti-Casein Kinase 1 delta/CSNK1D antibody [AF12G4]
See all Casein Kinase 1 delta/CSNK1D primary antibodies
DescriptionMouse monoclonal [AF12G4] to Casein Kinase 1 delta/CSNK1D
Tested applicationsSuitable for: IHC-P, Flow Cyt, WB, IPmore details
Species reactivityReacts with: Mouse, Rat, Human
Recombinant full length protein (His-tag) corresponding to Human Casein Kinase 1 delta/CSNK1D aa 1-415. Purified from E.coli.
Database link: P48730
- WB: HeLa, K562, SK-N-MC, 293T and L929 cell lysates. IHC-P: Human Alzheimer brain (hippocampus) tissue section.
This product was changed from ascites to tissue culture supernatant on 18th September 2017. Lot numbers higher than GR155202 will be from tissue culture supernatant. Please note that the dilutions may need to be adjusted accordingly.
Previously labelled as Casein Kinase 1 delta.
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In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
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Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferPreservative: 0.03% Sodium azide
Constituents: 0.01% BSA, 50% Glycerol, 0.87% Sodium chloride, HEPES
Concentration information loading...
PurityProtein G purified
Light chain typekappa
KO cell lines
KO cell lysates
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.
|IHC-P||Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|Flow Cyt||Use at an assay dependent concentration.
ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.
|WB||Use a concentration of 0.05 µg/ml. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).|
|IP||Use at an assay dependent concentration.
Use at 2μl
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.
modificationsAutophosphorylated on serine and threonine residues.
Cellular localizationCytoplasm. Nucleus.
- Information by UniProt
- Casein Kinase 1 delta antibody
- Casein kinase I delta isoform antibody
- Casein kinase I isoform delta antibody
All lanes : Anti-Casein Kinase 1 delta/CSNK1D antibody [AF12G4] (ab85320) at 1/500 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : CSNK1D knockout HeLa cell lysate
Lane 3 : K-562 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 47 kDa
Observed band size: 47 kDa
Lanes 1-3: Merged signal (red and green). Green - ab85320 observed at 47 kDa. Red - loading control, ab181602 observed at 37 kDa.
ab85320 Anti-Casein Kinase 1 delta/CSNK1D antibody [AF12G4] was shown to specifically react with Casein Kinase 1 delta/CSNK1D in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264667 (knockout cell lysate ab258383) was used. Wild-type and Casein Kinase 1 delta/CSNK1D knockout samples were subjected to SDS-PAGE. ab85320 and Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
All lanes : Anti-Casein Kinase 1 delta/CSNK1D 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/CSNK1D was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Mouse monoclonal to Casein Kinase 1 delta / CSNK1D 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/CSNK1D; non specific - 40kDa: We are unsure as to the identity of this extra band.
ab85320 has been referenced in 7 publications.
- Yamaguchi Y et al. Phosphorylated NF-?B subunit p65 aggregates in granulovacuolar degeneration and neurites in neurodegenerative diseases with tauopathy. Neurosci Lett 704:229-235 (2019). PubMed: 30946927
- Beale AD et al. Casein Kinase 1 Underlies Temperature Compensation of Circadian Rhythms in Human Red Blood Cells. J Biol Rhythms 34:144-153 (2019). PubMed: 30898060
- Shen S et al. Induction of Huh-7 cell apoptosis by HCV core proteins via CK1a-p53-Bid signaling pathway. Mol Med Rep 17:7559-7566 (2018). PubMed: 29620268
- Eng GWL et al. Site-specific phosphorylation of casein kinase 1 d (CK1d) regulates its activity towards the circadian regulator PER2. PLoS One 12:e0177834 (2017). PubMed: 28545154
- Wang SK et al. Fam83h null mice support a neomorphic mechanism for human ADHCAI. Mol Genet Genomic Med 4:46-67 (2016). WB . PubMed: 26788537
- Nishikawa T et al. The identification of raft-derived tau-associated vesicles that are incorporated into immature tangles and paired helical filaments. Neuropathol Appl Neurobiol 42:639-653 (2016). PubMed: 26501932
- Sinnberg T et al. Casein kinase 1a has a non-redundant and dominant role within the CK1 family in melanoma progression. BMC Cancer 16:594 (2016). IF . PubMed: 27488834