I have used Diagenode's antibodies for my ChIP–qPCR experiments in HK-2 cells. The antibodies performed very well in our experiments with specific signal, and good signal to noise ratio in the promoter of the gapdh gene.
Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K4me3 ChIP assays were performed using human HK-2 cells, the Diagenode antibody against H3K4me3 (Cat. No. C15410003) and optimized PCR primer pairs for qPCR. ChIP was performed with standard fixation and sonication protocol, using sheared chromatin from 4 million cells. A total amount of 4 ug of antibody were used per ChIP experiment, normal mouse IgG (4 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the active genes GAPDH and. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). These results are in accordance with the observation that trimethylation of K4 at histone H3 is associated with the promoters of active genes.
Claudio Cappelli PhD. Post-Doc. Investigator, Laboratory of Molecular Pathology, Institute of Biochemestry and Microbiology, University Austral of Chile, about H3K4me3 polyclonal antibody Premium, 50 μl size.
about H3K4me3 Antibody, H3K4me3 polyclonal antibody (sample size).
I used ChIP-qPCR with the Diagenode CRISPR/Cas9 polyclonal antibody to successfully show that Cas9 binds to the target region of my sgRNA, validating my CRISPR experiment. The antibody produced minimal background signal at non-specific genomic regions. I am now using the antibody to validate further sgRNA in different CRISPR cell lines.
ChIP was performed on Jurkat cells expressing dCas9-VP64-mCherry and a sgRNA targeting the IL1RN promoter. Each IP was performed using 4 million cells and 2 µL CRISPR/Cas9 polyclonal antibody (Diagenode C15310258) or 1 µg rabbit IgG control antibody (Diagenode C15410206). qPCR was carried out on undiluted ChIP DNA using SYBR green and PCR primers directed against the sgRNA binding site at IL1RN, as well as two non-target regions at the SLC4A1 and TP53 promoters. ChIP enrichment was measured using the percent input method.
Researcher from the University of Manchester
about CRISPR/Cas9 Antibody, CRISPR/Cas9 Antibody (sample size).
Diagenode’s CRISPR/Cas9 polyclonal antibody shows superior signal than the original clone 7A9: a researcher from EPFL in Lausanne, Switzerland has compared these two antibodies in Western blot.
Western blot was performed using HCT116 DKO cells transduced with Krab-dCas9 (2) or non-transduced (1) cells. Then, 100,000 cells were lysed in sample buffer 2x and boiled 5 min at 95°C before loading in a 15% acrylamide gel. The same sample was loaded 3x in the same gel. The membrane was cut in 3 parts for each antibody. Membrane was blocked 1h with 3% BSA at RT. Antibodies were diluted 1:1,000 in 3% BSA and incubated overnight at 4°C. Secondary incubation was done for 1h at RT (1:10,000 dilution). Anti-hnRNPA1 was used as a loading control.
EPFL in Lausanne
about CRISPR/Cas9 Antibody, CRISPR/Cas9 Antibody (sample size).
I have used Diagenode's antibodies for my ChIP Seq experiments. The antibodies performed very well in our experiments with specific signal, and good signal to noise ratio.
Antibodies used in our lab: Pol II Monoclonal Classic (C15200004).
Junaid Akhtar, Institute of Development Biology and Neurobiology, Mainz, Germany
about Pol II Antibody, Pol II Antibody (sample size).
I have been using Diagenode products to perform ChIP-seq during the last three years and I am very satisfied, with the Bioruptor, the kits and the antibodies. I have used the iDeal ChIP-seq kit for Histones and the iDeal ChIP-seq kit for Transcription Factors with very successful and reproducible results. Once I tried to ChIP histones with a home-made protocol and it worked much worse in comparison with Diagenode kits. In other occasion, I tried a non-Diagenode antibody for a transcription factor and I also got much poor results, however with the Diagenode antibody I always got very nice results. I strongly recommend the use of Diagenode products.
Dr. Francisca Martinez Real - Development and Disease Research Group - Max Planck Institute for Molecular Genetics, Berlin, Germany
about iDeal ChIP-seq kit for Histones, iDeal ChIP-seq kit for Histones, iDeal ChIP-seq kit for Transcription Factors, iDeal ChIP-seq kit for Transcription Factors, iDeal ChIP-seq Kit for Transcription Factors.
Diagenode Ago monoclonal antibody (Cat No. C15200167) immunoprecipitates the predicted miRNA target alleles very efficiently
Haruko Takeda, University of Liège, Belgium
about Ago (Argonautes) Antibody , Ago (Argonautes) Antibody , Ago (Argonautes) Antibody (sample size).
In life sciences, epigenetics is nowadays the most rapid developing field with new astonishing discoveries made every day. To keep pace with this field, we are in need of reliable tools to foster our research - tools Diagenode provides us with. From antibodies to automated solutions - all from one source and with robust support. Antibodies used in our lab: H3K27me3 polyclonal antibody – Premium, H3K4me3 polyclonal antibody – Premium, H3K9me3 polyclonal antibody – Premium, H3K4me1 polyclonal antibody – Premium, CTCF polyclonal antibody – Classic, Rabbit IgG.
Dr. Florian Uhle, Dept. of Anesthesiology, Heidelberg University Hospital, Germany
about H3K27me3 Antibody, H3K27me3 antibody (sample size), H3K4me3 Antibody, H3K4me3 polyclonal antibody (sample size), H3K9me3 Antibody, H3K4me1 Antibody (sample size), H3K4me1 Antibody, CTCF Antibody , CTCF Antibody (sample size), Rabbit IgG, H3K9me3 Antibody (sample size).
We use the Bioruptor sonication device and the antibody against the histone modification H3K4me3 in our lab. The Bioruptor device is used for the shearing of chromatin in ChIP-Seq experiments and for generation of protein lysates (whole cell extracts). Thanks to the Bioruptor device we achieve excellent and reproducible results in both applications. The H3K4me3 antibody is used in our ChIP-Seq experiments as well as Western Blotting. With this antibody we are able to generate highly enriched ChIP-Seq samples with extremely low background. In Western Blot we can detect one specific, strong band with the H3K4me3 antibody.
Diagenode provides not only very good products for research but also an excellent customer support.
Dr Lora Dimitrova, Charité-Universitätsmedizin Berlin, Germany
about Bioruptor® Plus sonication device, H3K4me3 Antibody, H3K4me3 polyclonal antibody (sample size).
I have been a Diagenode customer for over two years now. I have used the antibodies against the histone modifications H3K4me3, H3K4me2, H3K4me1, H3k27me3, H3K9ac, H3K27ac etc. provided by Diagenode to perform western blots.The high level of specificity of these antibodies in Pristionchus pacificus samples, confirmed by using several negative and positive controls run in parallel with synchronized culture samples, ensured successful and reproducible results.
Also, I am satisfied with the service and the support of personnel of Diagenode. They are prompt in replying to my emails and very helpful with their advice. Thanks a lot!
Vahan Serobyan, Prof. Dr Ralf J. Sommer's Lab, Max Planck Institute for Developmental Biology, Tübingen, Germany
I have extensively used the antibodies against the histone modifications H3K4me3, H3k27me3, H3K9ac, H4k8ac and H3K18ac provided by Diagenode. The high level of specificity and selectivity of these antibodies in mouse brain samples, confirmed by using several negative and positive controls run in parallel with mouse brain tissue samples, ensured successful and reproducible results. I have been a Diagenode costumer for over one year now and I am extremely satisfied with the efficiency of the Bioruptor Pico for chromatin shearing as well as all of the ChIP materials (i.e., antibodies, blocking peptides, primer pairs for qPCR) provided by this company. Many thanks.
Dr. Ermelinda Lomazzo, Institute of Physiological Chemistry, AG Prof. Beat Lutz. University Medical Center of the Johannes Gutenberg University Mainz, Germany
about H3K4me3 Antibody, H3K27me3 Antibody, H3K9ac Antibody, H4K8ac Antibody, H3K18ac Antibody, H3K27me3 Antibody (sample size).
I am working with the True MicroChIP & Microplex Library Preparation Kits and several histone modification antibodies like H3K27ac, H3K4me3, H3K36me3, and H3K27me3. I got always very good and reproducible results for my ChIP-seq experiments.
Andrea Thiesen, ZMB, Developmental Biology, Prof. Dr. Andrea Vortkamp´s lab, University Duisburg-Essen, Germany
about H3K27ac Antibody, H3K27ac Antibody (sample size), H3K4me3 Antibody, H3K4me3 polyclonal antibody (sample size), H3K36me3 Antibody, H3K36me3 Antibody (sample size), H3K27me3 Antibody, H3K27me3 antibody (sample size), True MicroChIP-seq Kit, MicroPlex Library Preparation Kit v2 (12 indexes).
I am working with Diagenode's antibodies for over 5 years now. The great and specific performance as well as the reliable high quality of the antibodies are crucial for the success of my ChIP experiments.
Dr Holger Bierhoff, Division Molecular Biology of the Cell II, Prof. Dr. Ingrid Grummt’s lab, DKFZ, Heidelberg, Germany
When doing CRISPR experiments, it is essential to verify the expression levels of the Cas9 to ensure high-quality results. Diagenode's new CRISPR/Cas9 4G10 antibody is extremely specific, able to recognize different Cas9 fusion proteins (even those used for CRISPRi experiments) and compatible with many biochemical assays, such as immunofluorescence, western blot and immunoprecipitation. Among the different antibodies in the market, Diagenode's 4G10 CRISPR/Cas9 is our antibody of choice.
Researcher at EPFL, Lausanne, Switzerland
about CRISPR/Cas9 Antibody 4G10 (sample size), CRISPR/Cas9 Antibody 4G10, CRISPR/Cas9 Antibody 4G10.
The antibody worked very well, even during our first attempt. We did a 1 hour incubation on a shaker at 4 degrees instead of overnight. We wanted to verify expression of Cas9 in various cell lines I had made (immunofluorescence). We have used the antibody a number of times and it works every time.
Researcher at Harvard University
about CRISPR/Cas9 Antibody 7A9, CRISPR/Cas9 Antibody 7A9 (sample size), CRISPR/Cas9 Antibody 7A9.
