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, IHC-P, IF-Cell |
| Clonality: | Monoclonal |
| Clone number: | SN06-50 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| 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, human tonsil tissue, human breast cancer tissue, HEK-293 cells treated with 100nM Calyculin A for 30 minutes. |
| Subcellular location: | Chromosome, Cytoplasm, Cytoskeleton, Nucleus. |
| Recommended Dilutions:
WB IHC-P IF-Cell |
1:1,000 1:200-1:1,000 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
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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. |
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Fig2:
Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-Phospho-Chk1 (S296) antibody (ET1611-76) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-76) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
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Fig3:
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-Phospho-Chk1 (S296) antibody (ET1611-76) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-76) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
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Fig4:
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”.