bsm-54050R
[Primary Antibody]
CLOCK (3F9) Monoclonal Antibody
DATASHEET
Host: Rabbit
Target Protein: CLOCK
Clonality: Monoclonal
Isotype: IgG
Entrez Gene: 9575
Swiss Prot: O15516
Source: Recombinant protein
Purification: Purified by Protein A.
Storage Buffer: 0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage: Store at 4°C for up to 2 weeks. For long term storage, store at -20°C in small aliquots to prevent freeze-thaw cycles.
Background:
Biological timepieces called circadian clocks are responsible for the regulation of hormonal rhythms, sleep cycles and other behaviors. The superchiasmatic nucleus (SCN), which is located in the brain, was the first mammalian circadian clock to be discovered. Clock, a member of the Basic-helix-loop-helix-psp (bHLH-PAS) family of transcription factors, has also been identified as having circadian function. Mutations within the clock gene have been shown to increase the length of the endogenous period and To contain a loss of rhythmicity of circadian oscillations. Clock contains a DNA-binding domain, a protein dimerization domain and a glutamine-rich C-terminal region, which indicates transactivation ability. It has been speculated that Clock may regulation circadian rhythmicity in combination with Other proteins such as Per. Per is also a PAS-domain containing protein that exhibits circadian function. Highest expression of Clock is seen in the hypothalamus and the eye.
Size: 100ul
Concentration: 1ug/ul
Predicted Molecular Weight: 95
Cross Reactive Species:
Human
Mouse
Rat
For research use only. Not intended for diagnostic or therapeutic use.
VALIDATION IMAGES
Lane 1: MCF-7 Cells; Lane 2: PC-12 Cells; Probed with CLOCK (3F9) Monoclonal Antibody (bsm-54050R) at 1:1000 overnight at 4°C followed by a conjugated secondary antibody for 60 minutes at 37°C.
IF(ICC) staining with CLOCK (3F9) Monoclonal Antibody (bsm-54050R) at 1:100 in Hela cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100/PBS.
IF(ICC) staining with CLOCK (3F9) Monoclonal Antibody (bsm-54050R) at 1:100 in SHG-44 cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100/PBS.
Paraformaldehyde-fixed and paraffin-embedded Human Fetal Skeletal Muscle tissue incubated with CLOCK (3F9) Monoclonal Antibody (bsm-54050R) at 1:100, overnight at 4°C, followed by a conjugated secondary antibody and DAB staining. Counterstained with hematoxylin.
Paraformaldehyde-fixed and paraffin-embedded Mouse Colon tissue incubated with CLOCK (3F9) Monoclonal Antibody (bsm-54050R) at 1:100, overnight at 4°C, followed by a conjugated secondary antibody and DAB staining. Counterstained with hematoxylin.
Paraformaldehyde-fixed and paraffin-embedded Human Colon Cancer tissue incubated with CLOCK (3F9) Monoclonal Antibody (bsm-54050R) at 1:100, overnight at 4°C, followed by a conjugated secondary antibody and DAB staining. Counterstained with hematoxylin.
Paraformaldehyde-fixed and paraffin-embedded Human Kidney tissue incubated with CLOCK (3F9) Monoclonal Antibody (bsm-54050R) at 1:100, overnight at 4°C, followed by a conjugated secondary antibody and DAB staining. Counterstained with hematoxylin.
ICC staining of CLOCK in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (bsm-54050R, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
Flow cytometric analysis of Hela cells with CLOCK (3F9) Monoclonal Antibody (bsm-54050R) at 1:100 dilution (red) compared with an unlabeled control (cells without incubation with primary antibody; black).
ICC staining of CLOCK in SHG-44 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (bsm-54050R, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
Flow cytometric analysis of CLOCK was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (bsm-54050R, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Paraformaldehyde-fixed, paraffin embedded (human colon); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (CLOCK (3F9)) Monoclonal Antibody, Unconjugated (bsm-54050R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023)instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human colon carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (CLOCK (3F9)) Monoclonal Antibody, Unconjugated (bsm-54050R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023)instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human fetal skeletal); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (CLOCK (3F9)) Monoclonal Antibody, Unconjugated (bsm-54050R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023)instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (CLOCK (3F9)) Monoclonal Antibody, Unconjugated (bsm-54050R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023)instructionsand DAB staining.