Casein Kinase 1 alpha Recombinant Rabbit Monoclonal Antibody [PSH04-46]
cat.: HA722131
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, FC, IP
Clonality: Monoclonal
Clone number: PSH04-46
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 39 kDa
Isotype: IgG
Immunogen: Recombinant protein within human CSNK1A1 aa 1-337 / 337.
Positive control: HCT 116 cell lysate, HeLa cell lysate, HEK-293 cell lysate, MCF7 cell lysate, Jurkat cell lysate, NIH/3T3 cell lysate, RAW264.7 cell lysate, PC-12 cell lysate, mouse brain tissue lysate, rat brain tissue lysate, HCT 116.
Subcellular location: Cytoplasm, cytoskeleton, microtubule organizing center, centrosome, Chromosome, centromere, kinetochore, Nucleus speckle, cilium basal body, spindle.
Recommended Dilutions:
  WB
  IF-Cell
  FC
  IP
1:2,000
1:500
1:1,000
1-2μg/sample
Uniprot #: SwissProt: P48729 Human | Q8BK63 Mouse | P97633 Rat
Alternative names: Casein kinase 1 alpha 1 Casein kinase I isoform alpha CK1 CK1A CKI alpha CKI-alpha CKIa Clock regulator kinase Csnk1a1 Down regulated in lung cancer HLCDGP1 KC1A_HUMAN PRO2975
Images
HA722131_1.jpg Fig1: Western blot analysis of Casein Kinase 1 alpha on different lysates with Rabbit anti-Casein Kinase 1 alpha antibody (HA722131) at 1/2,000 dilution and competitor's antibody 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 kDa
Observed band size: 34 kDa

Exposure time: 15 seconds; ECL: K1801;
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 (HA722131) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA722131_2.jpg Fig2: Immunocytochemistry analysis of HCT 116 cells labeling Casein Kinase 1 alpha with Rabbit anti-Casein Kinase 1 alpha antibody (HA722131) 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 (HA722131) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA722131_3.jpg Fig3: Flow cytometric analysis of HCT 116 cells labeling Casein Kinase 1 alpha.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA722131, 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 (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
HA722131_4.jpg Fig4: Casein Kinase 1 alpha was immunoprecipitated from 0.2 mg HeLa cell lysate with HA722131 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using HA722131 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: HeLa cell lysate (input)
Lane 2: HA722131 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of HA722131 in HeLa cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 1 minute 11 seconds; ECL: K1801
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.