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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H | ChIP, ICC, IH(P), WB | Rb | Affinity Purified | Polyclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | 07-596 |
| Brand Family | Upstate |
| Trade Name |
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| Description | Anti-Histone H4 (citrulline 3) Antibody |
| Alternate Names |
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| Background Information | Histone H4 (UniProt P62805; also known as H4 histone family member A, Histone 1 H4a) is encoded by the HIST1H4A (also known as H4/A, H4FA) gene (Gene ID: 8359) in human. Histone H4 is one of the main histone proteins that contributes to the chromatin structure in both prokaryotic and eukaryotic cells. Histone H4 can undergo several different types of epigenetic modifications that influence cellular processes, including methylation and acetylation on lysine and arginine residues, phosphorylation on serine and threonine residues, as well as conversion Arg3 to a citrulline residue by the peptidylarginine deiminase PAD4-catalyzed deimination reaction (citrullination). Histone H4 Arg3 citrullination (H4Cit3) weakens histone-DNA interaction as a result of a conformational change and a reduced positive charge at the histone H4 N-terminal end. Bacterial infection is known to stimulate H4Cit3 in neutrophils, forming highly decondensed extracellular chromatin structure NET (neutrophil extracellular trap) that could capture and kill microorganisms. On the other hand, citrullinated proteins are recognized as non-self proteins and can activate the immune system. A functional haplotype of the PADI4 gene has been shown to be associated with the pathogenesis of rheumatoid arthritis through induction of the antigenicity of proteins. H4Cit3 is induced in response to DNA damage through the p53-PADI4 pathway, resulting in upregulated H4Cit3 immunoreactivity localized around fragmented nuclei in apoptotic cells. In addition, H4Cit3 level is negatively correlated with p53 protein expression and tumor size in non-small cell lung cancer (NSCLC) tissues, indicating a crucial role of H4Cit3 in carcinogenesis. |
| Product Information | |
|---|---|
| Format | Affinity Purified |
| Presentation | Purified rabbit polyclonal antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Anti-Histone H4 (Citrulline 3), Cat. No. 07-596, is rabbit polyclonal antibody that detects Histone H4 citrullinated on Arginine 3 and is tested in ChIP, Immunocytochemistry, Immunohistochemistry (Paraffin), Peptide Inhibition Assay & Western Blotting. |
| Key Applications |
|
| Application Notes | Tested Applications Immunocytochemistry Analysis (ICC): A 1:500 dilution from a representative lot detected enhanced histone H4 Arg3 citrullination in ionomycin-stimulated primary human neutrophils. (Courtesy of Kerri Mowen, Ph.D., Padlock Therapeutics). Immunohistochemistry Analysis (IHC): A 1:250 dilution from a representative lot detected enhanced histone H4 Arg3 citrullination (H4Cit3) in 4% paraformaldehyde-fixed, decalcified, paraffin-embedded hind paw sections from wild-type, but not Pad4-knockout mice 5 days post immunization with arthritogenic serum from K/BxN mice (Courtesy of Kerri Mowen, Ph.D., Padlock Therapeutics). Peptide Inhibition Assay: Target band detection in acid extract of DMSO differentiated HL60 cells treated with calcium ionophore A23187 was prevented by preblocking of a representative lot with Histone H4 (citrulline 3) peptide. Chromatin Immunoprecipitation (ChIP) Analysis (ChIP): 1 µg from a representative lot immunoprecipitated Histone H4 (Citrulline 3) in chromatin from MCF-7 cells. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| Biological Information | |
|---|---|
| Immunogen | KLH-conjugated linear peptide corresponding to the first 10 amino acids of human histone H4 (Met1 removed), where Arginine 3 is citrullinated. C-terminal cysteine was added for conjugation purposes. |
| Epitope | N-terminus |
| Concentration | Please refer to lot specific datasheet. |
| Host | Rabbit |
| Specificity | This rabbit polyclonal antibody specifically detects Histone H4 citrullinated on Arginine 3. |
| Species Reactivity |
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| Species Reactivity Note | Human. Predicted to react with Mouse based on 100% sequence homology. |
| Antibody Type | Polyclonal Antibody |
| Entrez Gene Number |
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| Gene Symbol |
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| Modifications |
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| Purification Method | Affinity purified. |
| UniProt Number |
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| Molecular Weight | ~12 kDa observed; 11.37 kDa calculated. Uncharacterized bands may be observed in some lysate(s). |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Evaluated by Western Blotting in lysate from HL60 cells induced by retinoic acid and treated with calcium Ionophore (A23187). Western Blotting Analysis (WB): A 1:2,000 dilution of this antibody detected Histone H4 citrullinated on arginine 3 in lysates from HL60 cells induced by retinoic acid and either untreated (Lane 1) or treated with calcium ionophore (A23187) (Lane 2). |
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Recommend storage at +2°C to +8°C. For long term storage antibodies can be kept at -20°C. Avoid repeated freeze-thaws. |
| Packaging Information | |
|---|---|
| Material Size | 200 µL |