Casein Kinase 1 alpha Antibody (PSH04-46)

Immunoprecipitation: Casein Kinase 1 alpha Antibody (PSH04-46) [NBP3-32107] - Casein Kinase 1 alpha was immunoprecipitated from 0.2 mg HeLa cell lysate with NBP3-32107 at 2 ug/25 ul agarose. Western blot was performed from the immunoprecipitate using NBP3-32107 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input) Lane 2: NBP3-32107 IP in HeLa cell lysate Lane 3: Rabbit IgG instead of NBP3-32107 in HeLa cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 1 minute 11 seconds; ECL

Immunocytochemistry/ Immunofluorescence: Casein Kinase 1 alpha Antibody (PSH04-46) [NBP3-32107] - Immunocytochemistry analysis of HCT 116 cells labeling Casein Kinase 1 alpha with Rabbit anti-Casein Kinase 1 alpha antibody (NBP3-32107) at 1/500 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Casein Kinase 1 alpha antibody (NBP3-32107) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.

Flow Cytometry: Casein Kinase 1 alpha Antibody (PSH04-46) [NBP3-32107] - Flow cytometric analysis of HCT 116 cells labeling Casein Kinase 1 alpha. Cells were fixed and permeabilized. Then stained with the primary antibody (NBP3-32107, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Western Blot: Casein Kinase 1 alpha Antibody (PSH04-46) [NBP3-32107] - Western blot analysis of Casein Kinase 1 alpha on different lysates with Rabbit anti-Casein Kinase 1 alpha antibody (NBP3-32107) at 1/2,000 dilution.Lane 1: HCT 116 cell lysate (20 µg/Lane)Lane 2: HeLa cell lysate (20 µg/Lane)Lane 3: HEK-293 cell lysate (20 µg/Lane)Lane 4: MCF7 cell lysate (20 µg/Lane)Lane 5: Jurkat cell lysate (20 µg/Lane)Lane 6: NIH/3T3 cell lysate (20 µg/Lane)Lane 7: RAW264.7 cell lysate (20 µg/Lane)Lane 8: PC-12 cell lysate (20 µg/Lane)Lane 9: Mouse brain tissue lysate (40 µg/Lane)Lane 10: Rat brain tissue lysate (40 µg/Lane)Predicted band size: 39 kDaObserved band size: 34 kDaExposure time: 15 seconds; ECL;4-20% SDS-PAGE gel.Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.

• Reactivity: Hu, Mu, Rt
• Applications: WB, Flow, ICC/IF, IP
• Clone: PSH04-46
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

Casein Kinase 1 alpha Antibody (PSH04-46) Summary

• Description: Novus Biologicals Rabbit Casein Kinase 1 alpha Antibody (PSH04-46) (NBP3-32107) is a recombinant monoclonal antibody validated for use in WB, Flow, ICC/IF and IP. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Additional Information: Recombinant Monoclonal Antibody
• Immunogen: Recombinant protein within human Casein Kinase 1 alpha aa 1-337 / 337. (Uniprot: P48729)
• Localization: Cytoplasm, cytoskeleton, microtubule organizing center, centrosome, Chromosome, centromere, kinetochore, Nucleus speckle, cilium basal body, spindle.
• Source: HEK293
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Gene: CSNK1A1
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 1:1000
  • Immunocytochemistry/ Immunofluorescence 1:500
  • Immunoprecipitation 1-2ug/sample
  • Western Blot 1:2000
• Theoretical MW: 39 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS (pH7.4), 0.1% BSA and 40% Glycerol
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein A purified

Alternate Names for Casein Kinase 1 alpha Antibody (PSH04-46)

• Casein Kinase 1 alpha
• casein kinase 1, alpha 1
• casein kinase I isoform alpha
• CK1CKI-alpha
• CSNK1A1
• down-regulated in lung cancer
• EC 2.7.11.1
• HLCDGP1
• PRO2975

Background

Casein kinase I (also designated CKI) and casein kinase II (CKII) compose a family of serine/threonine protein kinases which are present in all eukaryotes examined to date. Casein kinase I family members, which include casein kinase Ialpha, Igamma, Idelta and Iepisilon, have been implicated in the control of cytoplasmic and nuclear processes, including DNA replication and repair. CKII is usually expressed as a tetrameric complex consisting of either an alpha2beta2 or an alphaalpha'beta2 structure. The a catalytic subunit is stimulated by the beta regulatory subunit, which undergoes autophosphorylation. Casein kinase II activity is high in the cytosol and nucleus of proliferating and differentiating cells. Casein kinase II is known to phosphorylate more than 100 different substrates including nuclear oncoproteins, transcription factors and enzymes involved in DNA metabolism.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: CSNK1A1