Overview

  • Product name

  • Description

    Rabbit polyclonal to Chk1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WB, ELISA, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • 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

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

This product has been referenced in:

  • Chen J  et al. The Ataxia telangiectasia-mutated and Rad3-related protein kinase regulates cellular hydrogen sulfide concentrations. DNA Repair (Amst) 73:55-63 (2019). Read more (PubMed: 30470507) »
  • Dias MH  et al. Fibroblast Growth Factor 2 lethally sensitizes cancer cells to stress-targeted therapeutic inhibitors. Mol Oncol 13:290-306 (2019). Read more (PubMed: 30422399) »
See all 19 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

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

Dr. Richelle Sopko

Verified customer

Submitted Aug 01 2013

Answer

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 has caused. As requested, I have issued a free of charge replacement with the order number 1212037 (ab47574).

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

Answer

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

Read More

Answer

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 send you a different replacement antibody against the same target. Could you confirm the catalogue number of antibody?

I will look forward to hearing from you soon.

Read More

Answer

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;

- Which cell line was used?
- Did you add miRNA against chk1 protein?
- Have you tried no primary control and loading control?

Thank you very much for your cooperation. I will look forward to hearing from you soon.

Read More
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

Abcam user community

Verified customer

Submitted Oct 31 2012

Answer

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 un remboursement ou un remplacement gratuit. Ces anticorps sonttestés en IHC-P et sur du tissue humain et de souris. Donc vous êtes couvert par notre garantie pour cette utilisation. Si néanmoins vous avez besoin d'utiliserces anticorps surdu tissue d'une autre espèce, veuillez me contacter.

Nous vous invitons aussi a utiliser notre système d'Abreviews. Notre système d'Abreviews permet aux chercheurs de soumettre leur étude sur ce produit en échange de Abpoints. Les Abpoints peuvent être échangés en remise sur un prochain achat ou récompensés par des achats sur Amazon.fr. En général, les récompensations pour les Abreviews sont assez généreuses:
https://www.abcam.com/abreviews

xx

J'espère ces informations vous sont utiles. N'hésitez pas à me contacter si vous avez besoin de plus de renseignements.

Read More
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

Dr. Ravi Dashnamoorthy

Verified customer

Submitted Mar 24 2008

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