Phospho-Chk1 (S345) Recombinant Rabbit Monoclonal Antibody [PS01-17]
cat.: HA721189
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse |
| Applications: | WB, IF-Cell |
| Clonality: | Monoclonal |
| Clone number: | PS01-17 |
| 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: | 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: 54 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phosphopeptide corresponding to residues surrounding Ser345 of human Chk1. |
| Positive control: | HEK-293 treated with 200nM Calyculin A for 1 hour cell lysate, NIH/3T3 treated with UV for 20 minutes then recover for 2 hours cell lysate, HeLa treated with Etoposide cell lysate, HeLa treated with Hydroxyurea cell lysate, HeLa treated with 4mM Hydroxyurea for 20 hours. |
| Subcellular location: | Nucleus, Chromosome, Cytoplasm, cytoskeleton, microtubule organizing center, centrosome. |
| Recommended Dilutions:
WB IF-Cell |
1:1,000 1:200 |
| Uniprot #: | SwissProt: O14757 Human | O35280 Mouse |
| Alternative names: | C85740 Cell cycle checkpoint kinase Checkpoint , S. pombe, homolog of, 1 Checkpoint kinase 1 Checkpoint kinase 1 homolog (S. pombe) CHEK 1 Chek1 Chk 1 Chk1 CHK1 checkpoint homolog (S. pombe) CHK1_HUMAN EC 2.7.11.1 rad27 Serine/threonine protein kinase Chk1 Serine/threonine-protein kinase CHK1 STT3, subunit of the oligosaccharyltransferase complex, homolog A (S. cerevisiae) |
Images
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Fig1:
Western blot analysis of Phospho-Chk1 (S345) on different lysates with Rabbit anti-Phospho-Chk1 (S345) antibody (HA721189) at 1/1,000 dilution. Lane 1: HEK-293 cell lysate Lane 2: HEK-293 treated with 200nM Calyculin A for 1 hour cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 54 kDa Observed band size: 56 kDa Exposure time: 59 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 (HA721189) at 1/1,000 dilution was used in primary antibody dilution (K1803) 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:
Western blot analysis of Phospho-Chk1 (S345) on different lysates with Rabbit anti-Phospho-Chk1 (S345) antibody (HA721189) at 1/1,000 dilution. Lane 1: NIH/3T3 cell lysate Lane 2: NIH/3T3 treated with UV for 20 minutes then recover for 2 hours cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 54 kDa Observed band size: 56 kDa Exposure time: 1 minute; 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 (HA721189) 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. |
|
Fig3:
Western blot analysis of Phospho-Chk1 (S345) on different lysates with Rabbit anti-Phospho-Chk1 (S345) antibody (HA721189) at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa treated with Etoposide cell lysate Lane 3: HeLa cell lysate Lane 4: HeLa treated with Hydroxyurea cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 54 kDa Observed band size: 56 kDa Exposure time: 5 minutes; 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 (HA721189) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature. |
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Fig4:
Immunocytochemistry analysis of HeLa cells treated with or without 4mM Hydroxyurea for 20 hours labeling Phospho-Chk1 (S345) with Rabbit anti-Phospho-Chk1 (S345) antibody (HA721189) at 1/200 dilution. Image shown an increased nuclear staining upon Hydroxyurea treatment. Cells were fixed in 4% paraformaldehyde for 20 minutes at 37 ℃, permeabilized with 0.1% Triton X-100 in PBS permeabilization for 5 minutes, and then blocked with 2% negative goat serum for 60 minutes at room temperature. Cells were then incubated with Rabbit anti-Phospho-Chk1 (S345) antibody (HA721189) at 1/200 dilution in 1% BSA 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. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.