H3pan Antibody 1B1B2
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.
| Lot | 003 |
|---|---|
| Concentration | 1.58 µg/µl |
| Species reactivity | Human, mouse, maize, tomato, poplar, Arabidopsis, barley: positive. Other species: not tested. |
| Type | Monoclonal, ChIP-grade |
| Purity | Protein A purified monoclonal antibody. |
| Host | Mouse |
| Storage Conditions | Store at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles. |
| Storage Buffer | PBS containing 0.05% azide. |
| Precautions | This 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.
- 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
Validation Data
Figure 1. ChIP results obtained with the monoclonal antibody directed against H3
ChIP assays were performed using human K562 cells, the 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).
Figure 2. Western blot analysis using the 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 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.
Figure 3. Western blot analysis using the monoclonal antibody directed against H3
Western blot was performed on whole cell extracts (30 µg) from different cell types (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 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.
Figure 4. Immunofluorescence using the monoclonal antibody directed against H3
HeLa cells were stained with the 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 labelled 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?
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
| WB 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 |
| IF 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... | Download |
| Epigenetic Antibodies Brochure BROCHURE More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e... | Download |
| Datasheet H3pan C15200011 DATASHEET Monoclonal antibody raised in mouse against histone H3, using a KLH-conjugated synthetic peptide ... | Download |
Safety sheets
Publications
How to properly cite our product/service in your workWe strongly recommend using this: H3pan Antibody 1B1B2 (Hologic Diagenode Cat# C15200011 Lot# 003). Click here to copy to clipboard. Using our products or services in your publication? Let us know! |
Conserved heat shock factors HvHSFA2 and HvHSFA3 control barley heat stress memory through diverged mechanisms |
Temporal modification of H3K9/14ac and H3K4me3 histone marksmediates mechano-responsive gene expression during the accommodationprocess in poplar |
Trichoderma root colonization triggers epigenetic changes in jasmonic andsalicylic acid pathway-related genes. |
Nox4 promotes endothelial differentiation through chromatin remodeling. |
Single amino-acid mutation in a Drosoph ila melanogaster ribosomalprotein: An insight in uL11 transcriptional activity. |
Activated Histone Acetyltransferase p300/CBP-Related SignallingPathways Mediate Up-Regulation of NADPH Oxidase,Inflammation, and Fibrosis in Diabetic Kidney |
ZNF354C is a transcriptional repressor that inhibits endothelialangiogenic sprouting. |
Revisiting promyelocytic leukemia protein targeting by human cytomegalovirus immediate-early protein 1. |
Widespread loss of the silencing epigenetic mark H3K9me3 in astrocytes and neurons along with hippocampal-dependent cognitive impairment in C9orf72 BAC transgenic mice. |
Class I TCP transcription factors target the gibberellin biosynthesis gene GA20ox1 and the growth promoting genes HBI1 and PRE6 during thermomorphogenic growth in Arabidopsis. |
Comprehensive Analysis of Chromatin States in Atypical Teratoid/Rhabdoid Tumor Identifies Diverging Roles for SWI/SNF and Polycomb in Gene Regulation. |
Epigenetic regulation of vascular NADPH oxidase expression and reactive oxygen species production by histone deacetylase-dependent mechanisms in experimental diabetes. |
The histone demethylase Phf2 acts as a molecular checkpoint to prevent NAFLD progression during obesity. |
Expression of the Parkinson's Disease-Associated Gene Alpha-Synuclein is Regulated by the Neuronal Cell Fate Determinant TRIM32 |