Anti-ChK1 (Checkpoint Kinase 1) Rabbit mAb

Catalog number :AT0388

Overview

Tested applications

ICC/IF	: Use a concentration of 5 µg/ml.
WB	: 1/500 - 1/1000. Detects a band of approximately 54 kDa (predicted molecular weight: 54 kDa).
ELISA	: 1/60000.
IHC-P	: Use a concentration of 4 µg/ml.

Properties

Immunogen: Synthetic non phosphopeptide derived from human Chk1 around the phosphorylation site of serine 345 (S-F-S^P-Q-P).

Form: Liquid
Preservative: 0.02% Sodium Azide
Constituents: 50% Glycerol, PBS, 150mM Sodium chloride, pH 7.4

Database links

Applications

WB Image: All lanes : Anti-Chk1 antibody (AT0388) 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

IHC Image: AT0388 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: Immunocytochemistry/ Immunofluorescence - Anti-Chk1 antibody

ICC/IF image of AT0388 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 AT0388 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit 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.