Phospho-Chk1 (S296) Recombinant Rabbit Monoclonal Antibody [SN06-50]
cat.: ET1611-76
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell
Clonality: Monoclonal
Clone number: SN06-50
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). 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: 54 kDa
Isotype: IgG
Immunogen: Synthetic phospho-peptide corresponding to residues surrounding Ser296 of human Chk1
Positive control: HEK-293 treated with 200nM Calyculin A for 1 hour cell lysate, NIH/3T3 treated with 100nM Calyculin A for 30 minutes cell lysate, C6 treated with 100ng/mL Calyculin A for 1 hour cell lysate, HEK-293 cells treated with 100nM Calyculin A for 30 minutes.
Subcellular location: Chromosome, Cytoplasm, Cytoskeleton, Nucleus.
Recommended Dilutions:
  WB
  IF-Cell

1:1,000
1:100
Uniprot #: SwissProt: O14757 Human | O35280 Mouse | Q91ZN7 Rat
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
ET1611-76_1.jpg Fig1: Western blot analysis of Phospho-Chk1 (S296) on different lysates with Rabbit anti-Phospho-Chk1 (S296) antibody (ET1611-76) 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
Lane 3: HEK-293 treated with 200nM Calyculin A for 1 hour cell lysate, then the membrane treated with λpp for 1 hour
Lane 4: NIH/3T3 cell lysate
Lane 5: NIH/3T3 treated with 100nM Calyculin A for 30 minutes cell lysate
Lane 6: NIH/3T3 treated with 100nM Calyculin A for 30 minutes cell lysate, then the membrane treated with λpp for 1 hour
Lane 7: C6 cell lysate
Lane 8: C6 treated with 100ng/mL Calyculin A for 1 hour cell lysate
Lane 9: C6 treated with 100ng/mL Calyculin A for 1 hour cell lysate, then the membrane treated with λpp for 1 hour

Lysates/proteins at 20 µg/Lane.

Predicted band size: 54 kDa
Observed band size: 54 kDa

Exposure time: Lane 1-3: 20 seconds; Lane 4-9: 3 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 (ET1611-76) 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.
ET1611-76_2.jpg Fig2: Immunocytochemistry analysis of HEK-293 cells treated with or without 100nM Calyculin A for 30 minutes labeling Phospho-Chk1 (S296) with Rabbit anti-Phospho-Chk1 (S296) antibody (ET1611-76) at 1/100 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-Chk1 (S296) antibody (ET1611-76) at 1/100 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.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.