Phospho-Rad17 (S656) Recombinant Rabbit Monoclonal Antibody [JE63-30]
cat.: HA722148
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse |
| Applications: | WB, IF-Cell, FC |
| Clonality: | Monoclonal |
| Clone number: | JE63-30 |
| Form: | Liquid |
| Storage condition: | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term. |
| 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: 77 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic peptide corresponding to residues surrounding Ser656 of human Rad17 protein. |
| Positive control: | HeLa treated with 100ng/mL Calyculin A for 30 minutes cell lysate, NIH/3T3 treated with 50ng/mL Calyculin A for 45 minutes cell lysate, HeLa cells treated with 100ng/mL Calyculin A for 30 minutes. |
| Subcellular location: | Nucleus. |
| Recommended Dilutions:
WB IF-Cell FC |
1:1,000 1:500 1:1,000 |
| Uniprot #: | SwissProt: O75943 Human | Q6NXW6 Mouse |
| Alternative names: | CCYC Cell cycle checkpoint protein Cell cycle checkpoint protein RAD17 FLJ41520 HRAD 17 hRad17 R24L Rad 17 Rad 24 RAD1 (S. pombe) homolog RAD1 homolog RAD17 RAD17 homolog (S. pombe) RAD17 homolog Rad17 like protein RAD17, S. pombe, homolog of RAD17_HUMAN RAD17Sp Rad24 Rad24, mouse, homolog of Rad24, S. cerevisiae, homolog of RF C activator 1 homolog RF C/activator 1 homolog RF-C/activator 1 homolog |
Images
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Fig1:
Western blot analysis of Phospho-Rad17 (S656) on different lysates with Rabbit anti-Phospho-Rad17 (S656) antibody (HA722148) at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa treated with 100ng/mL Calyculin A for 30 minutes cell lysate Lane 3: HeLa treated with 100ng/mL Calyculin A for 30 minutes cell lysate, then the membrane treated with λpp for 1 hour Lane 4: NIH/3T3 cell lysate Lane 5: NIH/3T3 treated with 50ng/mL Calyculin A for 45 minutes cell lysate Lane 6: NIH/3T3 treated with 50ng/mL Calyculin A for 45 minutes cell lysate, then the membrane treated with λpp for 1 hour Lysates/proteins at 20 µg/Lane. Predicted band size: 77 kDa Observed band size: 77 kDa Exposure time: 43 seconds; ECL: K1802; 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 (HA722148) 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. |
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Fig2:
Immunocytochemistry analysis of HeLa cells treated with or without 100ng/mL Calyculin A for 30 minutes labeling Phospho-Rad17 (S656) with Rabbit anti-Phospho-Rad17 (S656) antibody (HA722148) 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-Phospho-Rad17 (S656) antibody (HA722148) 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. |
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Fig3:
Flow cytometric analysis of HeLa cells treated with or without 100ng/mL Calyculin A for 30 minutes labeling Phospho-Rad17 (S656). Cells were fixed and permeabilized. Then stained with the primary antibody (HA722148, 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”.