Overview

  • Product name

    ChIP Kit - Plants
    See all ChIP Kit kits
  • Assay type

    Quantitative
  • Assay time

    6h 00m
  • Species reactivity

    Reacts with: Plants
  • Product overview

    Protein-DNA interactions play a critical role for cellular functions such as signal transduction, gene transcription and epigenetic silencing. In plants, interactions between the DNA-binding proteins and cognate promoter sequences are primary determinants in establishing spatial and temporal expression patterns of gene that affect homeostasis, development, and adaptation.


    Abcam's ChIP Kit - Plants (ab117137) offers an advantageous tool for identifying direct genome-wide associations between specific regulatory proteins and their target genes in plant cells within 6 hours. The kit is suitable for combining the specificity of immunoprecipitation with qualitative and quantitative PCR, MS-PCR, DNA sequencing and southern blot, as well as DNA microarray.


    This kit is designed for 24 or 48 ChIP reactions, not for 24 or 48 samples. The standard protocol of the kit allows for performing 8 reactions with one sample. For testing more samples, the amount of each sample should be reduced. The amount of each reagent used for chromatin preparation should be also proportionally reduced.

  • Notes

    ChIP assay products and guides

    Find more ChIP assay / chromatin immunoprecipitation resources and products, ChIP antibody products, and other ChIP assay kits and related reagents.

  • Tested applications

    Suitable for: ChIPmore details

Properties

  • Storage instructions

    Please refer to protocols.
  • Components 24 tests 48 tests
    100X Protease Inhibitor Cocktail 1 x 25µl 1 x 50µl
    5X Lysis Buffer I 1 x 12ml 1 x 24ml
    8-Well Assay Strips (with Frame) 3 units 6 units
    8-Well Strip Caps 3 units 6 units
    Antibody Buffer 1 x 15ml 1 x 30ml
    Anti-H3K9me2 (1 mg/mL) 1 x 5µl 1 x 8µl
    Binding Buffer 1 x 5ml 1 x 8ml
    ChIP Dilution Buffer 1 x 2ml 1 x 6ml
    DNA Release Buffer 1 x 2ml 2 x 2ml
    Elution Buffer 1 x 0.6ml 1 x 1.2ml
    F-Collection Tube 30 units 50 units
    F-Spin Column 30 units 50 units
    Lysis Buffer II 1 x 3ml 1 x 6ml
    Lysis Buffer III 1 x 2ml 1 x 4ml
    Lysis Buffer IV 1 x 1.5ml 1 x 5ml
    Normal Mouse IgG (1 mg/mL) 1 x 10µl 1 x 10µl
    Proteinase K (10 mg/mL) 1 x 25µl 1 x 50µl
    Reverse Buffer 1 x 2ml 2 x 2ml
    Wash Buffer 1 x 28ml 2 x 28ml
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab117137 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
ChIP Use at an assay dependent concentration.

Protocols

References

This product has been referenced in:

  • Baker K  et al. Chromatin state analysis of the barley epigenome reveals a higher order structure defined by H3K27me1 and H3K27me3 abundance. Plant J N/A:N/A (2015). Read more (PubMed: 26255869) »
See 1 Publication for this product

Customer reviews and Q&As

1-10 of 13 Abreviews or Q&A

Answer



I am happy to confirm that this kit is compatible with chromatin sheared using MNase (generally at the concentration of 5 Unit/ml for 5-10 min).

However, based on our testing, MNase digestion should be well monitored and controlled (for fragmenting DNA within 200-1000 bps).

Otherwise, it may cause over-degradation of DNA so that the DNA yield is lower than that obtained with sonication. We also recommended to perform the enzyme reaction in a small volume followed by heat inactivation of the enzyme then continuing the protocol at step 8 (tissue lysis and DNA shearing section, page 10) .

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Answer

1. Yes, the Covaris sonicator is fully comatible with this kit.
2. The Desi-Vac desiccator (vacuum pump container, YO-06525-02) from Cole -Parmer would be fine.

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Answer

Thank you for contacting us. The ChIP kit ab117137 should be suitable for testing with Chlamydomonas reinhardtii.

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

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Answer

Thank you for contacting us.

In general the chromatin extracts at the step of sonication should be clear. However in some of case, the green tinge may be seen to the chromatin extracts if the starting materials are with green color (ex: green
leaves). If the green color is seen in the pellet at each step including sonication, it could be caused by chlorophyll which is contained in the solution and may not be completely removed at each step. It is
also possible that tiny of chlorophyll is mixed in the pellet. However a green tinge to the chromatin extracts would not affect the chromatin immunoprecipitaion by antibody.

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

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https://www.abcam.com/abreviews

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Question
Answer

Could you also provide the Ct values for the EU-Hi region in both the IPd sample and input DNA?

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Answer

Thank you for contacting us.

I am sorry to hear you have been experiencing problems using our ChIP kit. In order to understand the problem in more detail would it be possible for you provide information to the following questions:

1. Total volume of diluted supernatant obtained at step 1 of " Protein/DNA immunoprecipitation"
2. Total volume of diluted supernatant used as "Input DNA"
3. Total volume of diluted supernatant used for sample IP
4. total volume of DNA elution for both sample and Input DNA
5. Total volume of both sample and Input DNA elution used for PCR.
6. Targeted gene by the primer pair that gives "high percentage" of input DNA.

Looking forward to your reply.

Read More

Answer

Thank you for your enquiry.

Regarding question 1:
You can reduce the volumes(and mass of plant tissue) accordingly to test moreplant samplesusing the samereagents provided with the kit. For example you can reduce the plant tissue by 10xand use 0.1g plant tissue per prep. Thiswould still produce sufficient chromatin for 1 ChIP.

Regarding question 2:
The "boiling" during fixation under vacum need not be a hard boil.Just a few bubbles produced is fine.

I hope this is helpful. Please contact me again if you have any further questions.

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Answer

Thank you very much for your reply.

The CTD repeat antibody, ab817, is expected to react with most species. We haven't specifically tested it against P. trichocarpa, however as long as the samples contain the CTD repeat I do expect it to react. The antibody does work in ChIPseq with samples from other species, so I also expect it to work with your samples.

I hope that this information will be useful, but please let me know if you have any further questions and I'll be happy to help.

Read More

Answer

Thank you very much for your call earlier this week and for your patience while I have been in touch with the lab regarding your enquiry.

The kit was tested with samples fromArabidopsisi thaliana in the lab, and based on user feedback and published articles, other plants (leaves and seeds)such as petunia have also been used. In principle, the kit should be suitable for use with root samples, though we do not have any reports of this being done in the past.

I hope that this information will be useful, but if you have any further questions please let me know and I'll be happy to help.

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Answer

Thank you very much for your email.

I have put in the attachment the list of the antibodies we have currently in our catalog which are tested against Arabidopsis.

I would also like to make you aware of a 100% Abreview testing discount we offer on untested applications and species. If you are interested in purchasing a product that is not tested in a particular species or application then we can issue a discount off a future purchase if the product is purchased and feedback is given to us via our Abreview system. We would offer the second primary antibody with a 100% discount. The Terms and Conditions of this offer can be found at: https://www.abcam.com/collaborationdiscount. If you have any further questions about this offer, then please do let me know.

Please let me know against which targets you are searching an antibody for, and I can see, what antibody we have or which antibody might be suitable for testing. Please mention also the applications in which you would need to use this antibodies.

I hope this information is helpful. Please do not hesitate to contact us again, could we assist you further.

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1-10 of 13 Abreviews or Q&A

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