CLOCK Recombinant Rabbit Monoclonal Antibody [JA12-33]
cat.: ET1704-82
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P, FC
Clonality: Monoclonal
Clone number: JA12-33
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 95 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human CLOCK aa 140-189 / 846.
Positive control: HeLa cell lysate, HEK-293 cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, human brain tissue lysate, human fetal skeletal tissue, human colon tissue, human colon carcinoma tissue, human kidney tissue, PC-12.
Subcellular location: Nucleus, Cytosol, Cytoplasm.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
1:500-1:5,000
1:50-1:200
1:50-1:200
1:50-1:200
1:50-1:100
Uniprot #: SwissProt: O15516 Human | O08785 Mouse | Q9WVS9 Rat
Alternative names: bHLHe8 Circadian locomoter output cycles kaput protein Circadian locomoter output cycles protein kaput Circadian Locomotor Output Cycles Kaput Circadium Locomotor Output Cycles Kaput Class E basic helix-loop-helix protein 8 CLOCK Clock circadian regulator Clock homolog Clock protein CLOCK_HUMAN hCLOCK KIAA0334
Images
ET1704-82_1.jpg Fig1: Western blot analysis of CLOCK on different lysates with Rabbit anti-CLOCK antibody (ET1704-82) at 1/1,000 dilution.

Lane 1: HeLa cell lysate (10 µg/Lane)
Lane 2: HEK-293 cell lysate (10 µg/Lane)
Lane 3: NIH/3T3 cell lysate (10 µg/Lane)
Lane 4: PC-12 cell lysate (10 µg/Lane)
Lane 5: Human brain tissue lysate (20 µg/Lane)

Predicted band size: 95 kDa
Observed band size: 95 kDa

Exposure time: 1 minute;

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 (ET1704-82) at 1/1,000 dilution was 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.
ET1704-82_2.jpg Fig2: Western blot analysis of CLOCK on different lysates with Rabbit anti-CLOCK antibody (ET1704-82) at 1/5,000 dilution.

Lane 1: HEK-293-si NT cell lysate
Lane 2: HEK-293-si CLOCK cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 95 kDa
Observed band size: 95 kDa

Exposure time: 50 seconds;

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 (ET1704-82) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/100,000 dilution was used for 1 hour at room temperature.
ET1704-82_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human fetal skeletal tissue using anti-CLOCK antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-82, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1704-82_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human colon tissue using anti-CLOCK antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-82, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1704-82_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-CLOCK antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-82, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1704-82_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-CLOCK antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-82, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1704-82_7.jpg Fig7: Immunocytochemistry analysis of PC-12 cells labeling CLOCK with Rabbit anti-CLOCK antibody (ET1704-82) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-CLOCK antibody (ET1704-82) at 1/100 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.
ET1704-82_8.jpg Fig8: Flow cytometric analysis of PC-12 cells labeling CLOCK.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1704-82, 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).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.