H3pan Antibody 1B1B2

Catalog Number
50 μg
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Monoclonal antibody raised in mouse against histone H3, using a KLH-conjugated synthetic peptide containing an unmodified sequence from the C-terminus of the protein. This antibody can be used as a loading control in both ChIP and WB experiments.

Concentration1.58 µg/µl
Species reactivityHuman, mouse, maize, tomato, poplar, arabidopsis: positive. Other species: not tested.
PurityProtein A purified monoclonal antibody.
Storage ConditionsStore at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles.
Storage BufferPBS containing 0.05% azide.
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP 1-2 µg/ChIP Fig 1
Western Blotting 1:1,000 - 1:5,000 Fig 2, 3
Immunofluorescence 1:500 Fig 4

* Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 µg per IP.

  • Validation Data

    H3pan Antibody ChIP Grade

    Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3
    ChIP assays were performed using human K562 cells, the Diagenode monoclonal antibody against H3 (cat. No. C15200011) and optimized PCR primer pairs for qPCR. ChIP was performed on sheared chromatin from 1 million cells. A titration consisting of 1, 2, 5 and 10 µg of antibody per ChIP experiment was analyzed. IgG (1 µg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoters of the active GAPDH and EIF4A2 genes, and for the inactive MYOD1 gene and the Sat2 satellite repeat. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

    H3pan Antibody validated in Western Blot

    Figure 2. Western blot analysis using the Diagenode monoclonal antibody directed against H3
    Western blot was performed on whole cell extracts from HeLa cells (40 µg, lane 1) and on 1 µg of recombinant histone H3 (lane 2) using the Diagenode monoclonal antibody against H3 (cat. No. C15200011). The antibody was diluted 1:2,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.

    H3pan Antibody validated in Western Blot

    Figure 3. Western blot analysis using the Diagenode monoclonal antibody directed against H3
    Western blot was performed on whole cell extracts (30 µg) from different celltypes (lane 1: HeLa, lane 2: K562, lane 3: MCF7, lane 4: U2OS, lane 5: HepG2, lane 6: Jurkat, lane 7: NIH3T3, lane 8: E14Tg2a mouse ES cells) using the Diagenode monoclonal antibody against H3 (Cat. No. C15200011). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.

    H3pan Antibody validated in Immunofluorescence

    Figure 4. Immunofluorescence using the Diagenode monoclonal antibody directed against H3
    HeLa cells were stained with the Diagenode monoclonal antibody against H3 (Cat. No. C15200011) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 1% BSA. The cells were immunofluorescently labeled with the H3 antibody (middle) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The left panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.

  • Loading Control

    H3pan monoclonal antibody can be used as a loading control for nuclear samples to compare the protein expression level between different samples.

    Why do you need a loading control?

    • To verify that equal sample amount was loaded on the gel
    • To confirm that the electrotransfer was done equally across the lines
    • To check that the signal detection after antibody incubation is homogenous across the lines

    What is the loading control?

    The loading control is an antibody against a protein which is highly expressed in certain types of samples. The level of protein stays constant in different conditions and in different organisms. The molecular weights of the protein of interest and loading control are different, allowing differentiation of both proteins as two different bands.

  • Target Description

    Histones are the main constituents of the protein part of chromosomes of eukaryotic cells. They are rich in the amino acids arginine and lysine and have been greatly conserved during evolution. Histones pack the DNA into tight masses of chromatin. Two core histones of each class H2A, H2B, H3 and H4 assemble and are wrapped by 146 base pairs of DNA to form one octameric nucleosome. Histones play a central role in the regulation of transcription, DNA repair, DNA replication and chromosomal stability. These different functions are established via a complex set of post-translational modifications which either directly or indirectly alter chromatin structure and DNA accessibility to facilitate transcriptional activation or repression or other nuclear processes.

  •  Applications
    Western blot : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies. Learn more about: Load... Read more
    ChIP-qPCR (ab)
    Read more
    Immunofluorescence: Diagenode offers huge selection of highly sensitive antibodies validated in IF. Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9 HeLa cells transfected with a Cas9 expression vector (... Read more
  •  Documents
    Antibodies you can trust POSTER
    Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar...
    Epigenetic Antibodies Brochure BROCHURE
    More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e...
    Datasheet H3pan C15200011 DATASHEET
    Monoclonal antibody raised in mouse against histone H3, using a KLH-conjugated synthetic peptide ...
  •  Safety sheets
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  •  Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: H3pan Antibody 1B1B2 (Diagenode Cat# C15200011 Lot# 003). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    Temporal modification of H3K9/14ac and H3K4me3 histone marksmediates mechano-responsive gene expression during the accommodationprocess in poplar
    Ghosh R. et al.
    Plants can attenuate their molecular response to repetitive mechanical stimulation as a function of their mechanical history. For instance, a single bending of stem is sufficient to attenuate the gene expression in poplar plants to the subsequent mechanical stimulation, and the state of desensitization can last for ...

    Trichoderma root colonization triggers epigenetic changes in jasmonic andsalicylic acid pathway-related genes.
    Agostini R. B. et al.
    Beneficial interactions between plant-roots and Trichoderma spp. lead to a local and systemic enhancement of the plant immune system through a mechanism known as priming of defenses. In recent reports, we outlined a repertoire of genes and proteins differentially regulated in distant tissues of maize plants previous...

    Nox4 promotes endothelial differentiation through chromatin remodeling.
    Hahner F. et al.
    RATIONALE: Nox4 is a constitutively active NADPH oxidase that constantly produces low levels of HO. Thereby, Nox4 contributes to cell homeostasis and long-term processes, such as differentiation. The high expression of Nox4 seen in endothelial cells contrasts with the low abundance of Nox4 in stem cells, which are a...

    Single amino-acid mutation in a Drosoph ila melanogaster ribosomalprotein: An insight in uL11 transcriptional activity.
    Grunchec H. et al.
    The ribosomal protein uL11 is located at the basis of the ribosome P-stalk and plays a paramount role in translational efficiency. In addition, no mutant for uL11 is available suggesting that this gene is haplo-insufficient as many other Ribosomal Protein Genes (RPGs). We have previously shown that overexpression of...

    Activated Histone Acetyltransferase p300/CBP-Related SignallingPathways Mediate Up-Regulation of NADPH Oxidase,Inflammation, and Fibrosis in Diabetic Kidney
    Alexandra-Gela Lazar et al.
    Accumulating evidence implicates the histone acetylation-based epigenetic mechanisms in the pathoetiology of diabetes-associated micro-/macrovascular complications. Diabetic kidney disease (DKD) is a progressive chronic inflammatory microvascular disorder ultimately leading to glomerulosclerosis and kidney failure. ...

    ZNF354C is a transcriptional repressor that inhibits endothelialangiogenic sprouting.
    Oo, James A and Irmer, Barnabas and Günther, Stefan and Warwick, Timothyand Pálfi, Katalin and Izquierdo Ponce, Judit and Teichmann, Tom andPflüger-Müller, Beatrice and Gilsbach, Ralf and Brandes, Ralf P andLeisegang, Matthias S
    Zinc finger proteins (ZNF) are a large group of transcription factors with diverse functions. We recently discovered that endothelial cells harbour a specific mechanism to limit the action of ZNF354C, whose function in endothelial cells is unknown. Given that ZNF354C has so far only been studied in bone and tum...

    Revisiting promyelocytic leukemia protein targeting by human cytomegalovirus immediate-early protein 1.
    Paulus C, Harwardt T, Walter B, Marxreiter A, Zenger M, Reuschel E, Nevels MM
    Promyelocytic leukemia (PML) bodies are nuclear organelles implicated in intrinsic and innate antiviral defense. The eponymous PML proteins, central to the self-organization of PML bodies, and other restriction factors found in these organelles are common targets of viral antagonism. The 72-kDa immediate-early prote...

    Widespread loss of the silencing epigenetic mark H3K9me3 in astrocytes and neurons along with hippocampal-dependent cognitive impairment in C9orf72 BAC transgenic mice.
    Jury N, Abarzua S, Diaz I, Guerra MV, Ampuero E, Cubillos P, Martinez P, Herrera-Soto A, Arredondo C, Rojas F, Manterola M, Rojas A, Montecino M, Varela-Nallar L, van Zundert B
    BACKGROUND: Hexanucleotide repeat expansions of the GC motif in a non-coding region of the C9ORF72 gene are the most common genetic cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Tissues from C9ALS/FTD patients and from mouse models of ALS show RNA foci, dipeptide-repeat proteins, an...

    Class I TCP transcription factors target the gibberellin biosynthesis gene GA20ox1 and the growth promoting genes HBI1 and PRE6 during thermomorphogenic growth in Arabidopsis.
    Ferrero LV, Viola IL, Ariel FD, Gonzalez DH
    Plants respond to a rise in ambient temperature by increasing the growth of petioles and hypocotyls. In this work, we show that Arabidopsis thaliana class I TEOSINTE BRANCHED 1, CYCLOIDEA, PCF (TCP) transcription factors TCP14 and TCP15 are required for optimal petiole and hypocotyl elongation under high ambient tem...

    Comprehensive Analysis of Chromatin States in Atypical Teratoid/Rhabdoid Tumor Identifies Diverging Roles for SWI/SNF and Polycomb in Gene Regulation.
    Erkek S, Johann PD, Finetti MA, Drosos Y, Chou HC, Zapatka M, Sturm D, Jones DTW, Korshunov A, Rhyzova M, Wolf S, Mallm JP, Beck K, Witt O, Kulozik AE, Frühwald MC, Northcott PA, Korbel JO, Lichter P, Eils R, Gajjar A, Roberts CWM, Williamson D, Hasselbla
    Biallelic inactivation of SMARCB1, encoding a member of the SWI/SNF chromatin remodeling complex, is the hallmark genetic aberration of atypical teratoid rhabdoid tumors (ATRT). Here, we report how loss of SMARCB1 affects the epigenome in these tumors. Using chromatin immunoprecipitation sequencing (ChIP-seq) on pri...

    Epigenetic regulation of vascular NADPH oxidase expression and reactive oxygen species production by histone deacetylase-dependent mechanisms in experimental diabetes.
    Manea SA, Antonescu ML, Fenyo IM, Raicu M, Simionescu M, Manea A
    Reactive oxygen species (ROS) generated by up-regulated NADPH oxidase (Nox) contribute to structural-functional alterations of the vascular wall in diabetes. Epigenetic mechanisms, such as histone acetylation, emerged as important regulators of gene expression in cardiovascular disorders. Since their role in diabete...

    The histone demethylase Phf2 acts as a molecular checkpoint to prevent NAFLD progression during obesity.
    Bricambert J, Alves-Guerra MC, Esteves P, Prip-Buus C, Bertrand-Michel J, Guillou H, Chang CJ, Vander Wal MN, Canonne-Hergaux F, Mathurin P, Raverdy V, Pattou F, Girard J, Postic C, Dentin R
    Aberrant histone methylation profile is reported to correlate with the development and progression of NAFLD during obesity. However, the identification of specific epigenetic modifiers involved in this process remains poorly understood. Here, we identify the histone demethylase Plant Homeodomain Finger 2 (Phf2) as a...

    Expression of the Parkinson's Disease-Associated Gene Alpha-Synuclein is Regulated by the Neuronal Cell Fate Determinant TRIM32
    Pavlou MA et al.
    Alpha-synuclein is an abundant neuronal protein which has been associated with physiological processes like synaptic function, neurogenesis, and neuronal differentiation but also with pathological neurodegeneration. Indeed, alpha-synuclein (snca) is one of the major genes implicated in Parkinson's disease (PD). Howe...

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