Anti-Chk1 antibody (ab47574)
- Product nameAnti-Chk1 antibodySee all Chk1 primary antibodies ...
- DescriptionRabbit polyclonal to Chk1
- Tested applicationsICC/IF, WB, ELISA, IHC-P more details
- Species reactivityReacts with: Mouse, Rat, Human, Fruit fly (Drosophila melanogaster)
Synthetic non phosphopeptide derived from human Chk1 around the phosphorylation site of serine 345 (S-F-SP-Q-P).
- Positive control
- HT29 cell extracts. This antibody gave a positive result in IF in the following Formaldehyde fixed cell line: MCF-7.
- Storage instructionsStore at -20°C. Stable for 12 months at -20°C
- Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 50% Glycerol, PBS, 150mM Sodium chloride, pH 7.4
- Concentration information loading...
- PurityImmunogen affinity purified
- Purification notesThe antibody was affinity purified from rabbit antiserum by affinity chromatography using epitope specific immunogen.
- Clonality Polyclonal
- Research Areas
Our Abpromise guarantee covers the use of ab47574 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||ICC/IF: Use a concentration of 5 µg/ml.|
|WB||WB: 1/500 - 1/1000. Detects a band of approximately 54 kDa (predicted molecular weight: 54 kDa).|
|IHC-P||IHC-P: Use a concentration of 4 µg/ml.|
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Anti-Chk1 antibody images
All lanes : Anti-Chk1 antibody (ab47574) at 1/500 dilution
Lane 1 : HT29 cell extract treated with UV. No peptide.
Lane 2 : HT29 cell extract treated with UV. Synthetic peptide present.
Predicted band size : 54 kDa
Observed band size : 54 kDa
ab47574 staining Chk1 in Human heart.
Left panel: with primary antibody at 4 ug/ml. Right panel: isotype control.
Sections were stained using an automated system (DAKO Autostainer Plus), at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers EDTA pH 9.0. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
ICC/IF image of ab47574 stained MCF-7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab47574 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
References for Anti-Chk1 antibody (ab47574)
This product has been referenced in:
- Fan S et al. DIM (3,3'-diindolylmethane) confers protection against ionizing radiation by a unique mechanism. Proc Natl Acad Sci U S A 110:18650-5 (2013). WB ; Rat . Read more (PubMed: 24127581) »
- Zhang J et al. NANOG modulates stemness in human colorectal cancer. Oncogene : (2012). Read more (PubMed: 23085761) »