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: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
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: 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
HA721189_1.jpg 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: 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;

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.
HA721189_2.jpg 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: 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.
HA721189_3.jpg Fig3: 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”.