Overview

  • Product nameAnti-Chk1 antibody
    See all Chk1 primary antibodies
  • Description
    Rabbit polyclonal to Chk1
  • Tested applicationsICC/IF, WB, ELISA, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Fruit fly (Drosophila melanogaster)
  • Immunogen

    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.

Properties

Applications

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.

Application Abreviews Notes
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.

Target

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

    Secondary antibody - anti-rabbit HRP (ab6721)

  • 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:
  • Gray JE  et al. A phase I, pharmacokinetic, and pharmacodynamic study of panobinostat, an HDAC inhibitor, combined with erlotinib in patients with advanced aerodigestive tract tumors. Clin Cancer Res 20:1644-55 (2014). IHC ; Human . Read more (PubMed: 24429877) »
  • Mashukova A  et al. The BAG-1 isoform BAG-1M regulates keratin-associated Hsp70 chaperoning of aPKC in intestinal cells during activation of inflammatory signaling. J Cell Sci 127:3568-77 (2014). Read more (PubMed: 24876225) »

See all 7 Publications for this product

Product Wall

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (4-20% gel)
Sample Fruit fly (Drosophila melanogaster) Tissue lysate - whole (0-4hr embryo)
Specification 0-4hr embryo
Treatment control shRNA and DChk1 (grp) shRNA embryos
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Dr. Richelle Sopko

Verified customer

Submitted Aug 01 2013

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this h...

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Could you let me know the purchase order number of ab47574 so that I can place an order for you?

Thank you for your email.

I am sorry from the details provided and with literature search I am unable to find a reason why the antibody is not giving the expected band size. In order to ensure, the problem is not due to antibody I am happy to ...

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Thank you for your email. I am sorry to hear that you have been experiencing problems with this antibody.

I have read the details you have kindly provided and have following further questions for better understanding of the problem;

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HeLa cell line)
Loading amount 40 µg
Specification HeLa cell line
Treatment 50 nM mitomycin C 6 hours
Gel Running Conditions Reduced Denaturing (8%)
Blocking step BSA as blocking agent for 45 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Oct 31 2012

Merci pour votre requête.

Nous n'avonsmalheureusementpas d'échantillons ou de petit volume de nos produits. Notre politique est que si un anticorps ne marche pas comme spécifié dans sa fiche technique, nous offrons...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Fruit fly (Drosophila melanogaster) Cell lysate - whole cell (Kc167)
Loading amount 10 µg
Specification Kc167
Gel Running Conditions Reduced Denaturing (10% SDS PAGE)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Ravi Dashnamoorthy

Verified customer

Submitted Mar 24 2008

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"