WB
Western blot analysis of Phospho-Chk1 (S345) on different lysates with Rabbit anti-Phospho-Chk1 (S345) antibody 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. 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 at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.ICC/IF
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 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 at 1/200 dilution in 1% BSA overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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.| Product Name | Phospho-Chk1 (S345) Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic phosphopeptide corresponding to residues surrounding Ser345 of human Chk1. |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ICC/IFWB |
| WB:1:1000 ICC/IF:1:200 |
|
| Species Reactivity | HumanMouse |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | CHEK1 |
|---|---|
| Gene Synonyms | CHK1 OZEMA21 |
| Gene Full Name | checkpoint kinase 1 |
| Gene Summary | The protein encoded by this gene belongs to the Ser/Thr protein kinase family. It is required for checkpoint mediated cell cycle arrest in response to DNA damage or the presence of unreplicated DNA. This protein acts to integrate signals from ATM and ATR, two cell cycle proteins involved in DNA damage responses, that also associate with chromatin in meiotic prophase I. Phosphorylation of CDC25A protein phosphatase by this protein is required for cells to delay cell cycle progression in response to double-strand DNA breaks. Several alternatively spliced transcript variants have been found for this gene. [provided by RefSeq, Oct 2011] |
| Molecular Weight(MW) | 54kDa(Observed band size:56kDa) |
| Function | Checkpoint kinase 1, commonly referred to as Chk1, is a serine/threonine-specific protein kinase that, in humans, is encoded by the CHEK1 gene. Chk1 coordinates the DNA damage response (DDR) and cell cycle checkpoint response. Activation of Chk1 results in the initiation of cell cycle checkpoints, cell cycle arrest, DNA repair and cell death to prevent damaged cells from progressing through the cell cycle. Checkpoint kinases (Chks) are protein kinases that are involved in cell cycle control. Two checkpoint kinase subtypes have been identified, Chk1 and Chk2. Chk1 is a central component of genome surveillance pathways and is a key regulator of the cell cycle and cell survival. Chk1 is required for the initiation of DNA damage checkpoints and has recently been shown to play a role in the normal (unperturbed) cell cycle. Chk1 impacts various stages of the cell cycle including the S phase, G2/M transition and M phase. In addition to mediating cell cycle checkpoints, Chk1 also contributes to DNA repair processes, gene transcription, egg production, embryo development, cellular responses to HIV infection and somatic cell viability. |
| Cellular Localization | Nucleus, Chromosome, Cytoplasm, cytoskeleton, microtubule organizing center, centrosome. |
WB
Western blot analysis of Phospho-Chk1 (S345) on different lysates with Rabbit anti-Phospho-Chk1 (S345) antibody 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. 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 at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
ICC/IF
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 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 at 1/200 dilution in 1% BSA overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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.| Application Notes | WB:1:1000 ICC/IF:1:200 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
Data sheet for OM644044