Overview

  • Product name

  • Description

    Rabbit polyclonal to p19 INK4d
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide corresponding to a region within N terminal amino acids 22-51 of Human p19 INK4d (UniProt ID: P55273) conjugated to KLH.

  • Positive control

    • Human testis lysate; testis tissue IF: HeLa cell line

Properties

Applications

Our Abpromise guarantee covers the use of ab102842 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
WB 1/100 - 1/500. Predicted molecular weight: 18 kDa.
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF Use a concentration of 5 µg/ml.

Target

  • Relevance

    Cyclins are important in regulating the cell cycle through their formation of enzymatic complexes with various cyclin dependent kinases. The activity of the cyclin D dependent kinases is controlled by inhibitors such as the INK4 family, which includes INK4a, INK4b, INK4c, and INK4d (p19 INK4d). p19 INK4d is a specific inhibitor of cdk 4 / cdk 6 and is periodically expressed during cell cycle. It is maximally induced as cells enter S phase.
  • Cellular localization

    Cytoplasmic and Nuclear
  • Database links

  • Alternative names

    • CDK inhibitor p19INK4d antibody
    • CDKN2D antibody
    • cell cycle inhibitor, Nur77 associating protein antibody
    • Cyclin dependent kinase inhibitor 2D antibody
    • Cyclin-dependent kinase 4 inhibitor D antibody
    • Cyclin-dependent kinase inhibitor 2D (p19, inhibits CDK4) antibody
    • inhibitor of cyclin-dependent kinase 4d antibody
    • INK4D antibody
    • MGC109660 antibody
    • MGC137547 antibody
    • MGC73010 antibody
    • p19 antibody
    • p19-INK4D antibody
    • p19INK4d antibody
    • Similar to cyclin-dependent kinase inhibitor 2D antibody
    see all

Images

  • Anti-p19 INK4d antibody (ab102842) at 1/100 dilution + Mouse testis tissue lysate at 35 µg

    Predicted band size: 18 kDa

  • ab102842, at a 1/50 dilution, staining P19 INK4d in formalin fixed, paraffin embedded testis tissue by Immunohistochemistry.
  • ICC/IF image of ab102842 stained HeLa cells. The cells were 4% paraformaldehdye 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 (ab102842, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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:

  • Prikrylova T  et al. 5-hydroxymethylcytosine Marks Mammalian Origins Acting as a Barrier to Replication. Sci Rep 9:11065 (2019). Read more (PubMed: 31363131) »
  • Rabachini T  et al. BOK promotes chemical-induced hepatocarcinogenesis in mice. Cell Death Differ 25:706-718 (2018). WB, IHC-P ; Mouse . Read more (PubMed: 29229991) »
See all 5 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Answer

Thank you for your reply.

I have processed your request to receive a new vial of ab102842 to try further dilutions of the antibody. Your new order number is * and you should receive that antibody tomorrow.

If there is anything else I can help you with, or if you have any issues with the new vial of the antibody, please let me know.

Read More

Answer

Thank you for your reply.

The recommended dilutions on the datasheets are only intended as a useful starting point for our customers, we strongly advise to optimize the antibody conditions for use in your own experiments.

I would be happy to either send you a new vial of ab102842 to continue troubleshooting this issue, or I can send a different antibody against the same target as a free of charge replacement. Please let me know how you would like to proceed and also please include either the order or PO number for this antibody so that I can track down your original order as quickly as possible.

I look forward to your reply and helping resolve this issue.

Read More

Answer

Thank you for your reply.

Based on the information that you have provided, I think that the following protocol suggestions may prove to be successful:

1 - Switch from a 5% milk to a 5% BSA block, this should help to cut down on background.

2 - Reduce your primary antibody concentration to 1/250, as this will also help reduce the level of background that you are seeing.

If neither of these variations prove to be successful, then please let me know and I will be happy to replace or refund the cost of the antibody for you.

I look forward to your reply and helping resolve this issue for you.

Read More

Answer

Thank you for your reply.

I would be very happy to try and help resolve this issue, if we cannot reduce the levels of background you are getting using ab102842, then I will be happy to send a replacement. To be able to help further in this matter, would you be able to answer a few protocol questions:

1 - What sample type are you using?

2 - Of the bands that you are seeing, are any at the predicted molecular weight?

3 - How much sample are you loading?

4 - What type of blocking agent are you using, milk, BSA etc?

5 -Apart from the 1/100, primary antibody concentration, have you tried anything else and how long do you incubate the sample for?

I look forward to your reply and helping to resolve this issue for you.

Read More

Question
Answer

Thank you for contacting us.

Yes, the antibody would still work if residues 47-51 are missing from the immunogen region.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Answer

Thank you for your response and for passing some useful details to me. RT-PCR data should be treated carefully since, there is no clear correlation between mRNA and protein levels and sometimes it is not easy to predict the protein expression levels from quantitative mRNA data. There are several publications available which delineate the technical boundaries of the approaches for quantitative analysis of protein expression and reveal that simple deduction from mRNA transcript analysis is insufficient. p19 INK4d is expressed at high levels in normal brain, testis, gastro-intestinal (digestive) tissues; and in different tumours (such as head and neck, glioma, cervix, stomach, colorectal,  melanoma, skin cancers). Since this antibody recognizes human and mouse p19 INK4d, any of these tissues can be applied as good positive control. I hope this helps and if I can assist further, please do not hesitate to contact me.      

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