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Histone H2a

Histone H2a is a core histone protein that plays a critical role in the organization and regulation of chromatin structure within eukaryotic cells.
It is one of the five main histone proteins responsible for packaging DNA into nucleosomes, the fundamental units of chromatin.
Histone H2a contributes to the stability and compaction of the nucleosome, influencing gene expression, DNA repair, and other essential cellular processes.
Reserach into Histone H2a is crucial for understanding chromatin dynamics and its impact on various biological functions.
PubCompare.ai's AI-driven platform offers a powerful tool to streamline and optimize Histone H2a research protocols, helpping scientiests effortelessly identify the best methodologies from literature, preprints, and patents.

Most cited protocols related to «Histone H2a»

1
Characterization of mTurquoise Fluorescent Protein
Cited 28 times
Expression of mTurquoise and its variants in E. coli was measured at 37 °C using a fluorescence microplate reader, as described9 (link), except that we used the pRSET plasmids for expression without induction. For brightness studies in mammalian cells, mTurquoise variants were excised from the pRSET vector using NheI and BsrGI and ligated into the CFP-2A-SYFP2 co-expression vector that was partially digested with NheI and BsrGI, and brightness analysis was performed as described10 (link) 2 days after transfection. For expression of mTurquoise variants in HeLa cells, we replaced the fluorescent protein from pSCFP3A-C1 with the mTurquoise variant using AgeI/BsrGI restriction sites. Photostability was measured in living HeLa cells by continuous illumination with light from a 100 W mercury lamp that was passed through a 436/20-nm excitation filter and reflected onto the sample with a 455DCLP dichroic mirror (Chroma). A ×63 oil immersion objective (Plan Apochromat NA 1.4) was used and a light intensity of 1.4–1.6 W cm−2 was determined when a ND1.3 grey filter was used, whereas a light intensity of 22–23 W cm−2 was determined without grey filter. To evaluate photostability under laser scanning conditions, we used HeLa cells expressing CFP variants tagged with histone 2A. A Nikon A1 laser scanning microscope was used and cells were imaged using a Nikon Plan Apo VC ×60 oil objective and a zoom factor of 6. Excitation was at 443 nm, and fluorescence was passed through a completely opened pinhole and a 482/35-nm bandpass filter. All parameters, including detector gain, laser intensity, and scan speed were identical between samples. Spectral imaging microscopy was performed on single cells as described10 (link).
Goedhart J., von Stetten D., Noirclerc-Savoye M., Lelimousin M., Joosen L., Hink M.A., van Weeren L., Gadella TW J.r, & Royant A. (2012). Structure-guided evolution of cyan fluorescent proteins towards a quantum yield of 93%. Nature Communications, 3, 751-.
Publication 2012
A-factor (Streptomyces) Cells Cloning Vectors Escherichia coli factor A Fluorescence HeLa Cells Histone H2a Laser Scanning Microscopy Light Mammals Mercury Microscopy Plasmids Proteins Radionuclide Imaging Submersion Transfection
2
Validation of Bovine Reference Genes
Cited 15 times
Eight reference genes were selected (ACTB, GAPD, Histone H2A, TBP, HPRT1, SDHA, YWHAZ and 18S rRNA) that belong to different functional classes to reduce the chance that the genes might be co-regulated (Table 3).
Primers for Histone H2A were taken from Robert and colleagues [19 (link)], primers for TBP were taken from Vigneault and colleagues [38 (link)] and primers for ACTB were taken from Fair and colleagues [27 (link)]. The other primers were designed by the Primer 3 software [39 (link)] and were based on RNA or DNA sequences found in Genbank. The reported bovine sequences were preferentially used and the specificity of the primers was tested using a BLAST analysis against the genomic NCBI database. PCR amplicons were characterized using Mfold [40 (link)] in order to predict the nature of any secondary structures which might influence the PCR efficiency. The PCR products were cloned (pCR 2.1 vector, Invitrogen, Merelbeke) and sequenced for verification (Thermo Sequenase Primer Cycle Sequencing Kit, Amersham Bioscience, Roosendaal) with a ALF Express sequencer (Amersham Bioscience, Roosendaal) [GenBank: DQ066891, DQ066892, DQ066893, DQ066894, DQ066895, DQ066896, DQ066897 and DQ066898]. Primer and amplicon information are listed in Table 1.
Goossens K., Van Poucke M., Van Soom A., Vandesompele J., Van Zeveren A, & Peelman L.J. (2005). Selection of reference genes for quantitative real-time PCR in bovine preimplantation embryos. BMC Developmental Biology, 5, 27.
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Publication 2005
Cattle Cloning Vectors Genes Genome Glyceraldehyde-3-Phosphate Dehydrogenases Histone H2a Oligonucleotide Primers RNA, Ribosomal, 18S thermo sequenase
3
Histone Octamer Refolding Protocol
Cited 11 times
Octamers containing desired histone compositions were assembled as previously described18 (link) with minor modifications. In brief, histones were dissolved in histone unfolding buffer (6 M guanidine hydrochloride, 20 mM Tris, 5 mM DTT, pH 7.5 at 4 °C) and combined in equimolar ratios (0.75 nmol each of desired version of histones H2A, H2B, H3, and H4). The total histone concentration was adjusted to 1 mg/ml, and the mixtures were placed in Slide-A-Lyzer MINI dialysis devices (3.5 kDa MW cutoff, ThermoFisher Scientific) and dialysed at 4 °C against 3 × 400 ml of octamer refolding buffer (2 M NaCl, 10 mM Tris, 0.5 mM EDTA, 1 mM DTT, pH 7.8 at 4 °C) for at least 4 h for each step, with one dialysis step overnight. The mixtures were then transferred to clean microcentrifuge tubes and spun down at 17,000g for 5 min at 4 °C to remove any precipitate. Supernatants were transferred to fresh microcentrifuge tubes, and 50% (v/v) glycerol was added. Octamer concentrations were measured by UV spectroscopy at 280 nm, and stored at −20 °C until use in nucleosome assembly.
Dann G.P., Liszczak G.P., Bagert J.D., Müller M.M., Nguyen U.T., Wojcik F., Brown Z.Z., Bos J., Panchenko T., Pihl R., Pollock S.B., Diehl K.L., Allis C.D, & Muir T.W. (2017). ISWI chromatin remodellers sense nucleosome modifications to determine substrate preference. Nature, 548(7669), 607-611.
Publication 2017
Buffers Dialysis Edetic Acid Glycerin Histone H2a Histones Hydrochloride, Guanidine Medical Devices Nucleosomes Sodium Chloride Spectrum Analysis Tromethamine
4
Immunofluorescence Staining of Chromatin
Cited 9 times
Coverslips with the fixed cells were removed from the plates and processed by floating on drops kept on hydrophobic laboratory film. After washing with PBS, the samples were permeabilized for 1′ with 0.5% Triton X100 in PBS, washed again with PBS and blocked for 20′ with blocking buffer (3% normal donkey serum, 3% cold water fish gelatin, 1% BSA, and 0.05% Tween 20 in PBS). Solutions of antibody (mouse mAB PL2-3) directed against the subnucleosomal complex of Histone 2A, Histone 2B, and chromatin (Losman et al., 1992 (link)) and against neutrophil elastase (rabbit pAB Calbiochem 481001) were applied in blocking buffer for 1–2 h. After washing with PBS, dye-conjugated secondary antibody solutions were applied for 30′–1 h (donkey anti mouse Cy3, donkey anti rabbit AlexaFluor 488, Jackson). The samples were washed with PBS and distilled water, stained with aqueous Hoechst 33342 (100 ng/ml, Sigma), washed again in distilled water and mounted with Mowiol.
Brinkmann V., Goosmann C., Kühn L.I, & Zychlinsky A. (2013). Automatic quantification of in vitro NET formation. Frontiers in Immunology, 3, 413.
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Publication 2012
Buffers Cells Chromatin Cold Temperature Equus asinus Fishes Gelatins Histone H2a Histones HOE 33342 Immunoglobulins Mus neutrophil elastase, human Rabbits Serum Triton X-100 Tween 20
5
Synthesis and Purification of Modified Histone Peptides
Cited 9 times

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Publication 2010
4-methylmorpholine acetonitrile Amino Acids Biotin Ethyl Ether High-Performance Liquid Chromatographies Histone H2a Histone H3 Ninhydrin Peptides piperidine Resins, Plant Rink amide resin Solvents Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Trifluoroacetic Acid

Most recents protocols related to «Histone H2a»

1
Antibody Validation for Histone Modifications
Antibody against anti-histone H2A.Z (1:5000; ab4626) was purchased from Abcam; antibody against Rpd3 (1:500; sc-514160) was purchased from Santa Cruz Biotechnology; antibodies against GFP (1:5000; 66002-1-1g), Myc (1:5000; 60003-2-1g), goat polyclonal anti-mouse IgG (1:5000; SA00001-1), and goat polyclonal anti-rabbit IgG (1:5000; SA00001-2) were obtained from Proteintech; antibody against pan–acetyl-lysine (1:2000, 9441S) was purchased from Cell Signaling Technology; antibody against FLAG M2 (1:3000; F1804-1MG) was obtained from Sigma-Aldrich; antibody against CBP (1:2000; Abs130593) was purchased from Absin Bioscience Inc.; and antibodies against Ino80 (1:500) and Ino80 K929ac (1:500) were custom-made in Abclonal. The specificity of the custom-made antibodies was confirmed by immunoblot analysis with cell extract or IP of corresponding mutants (Fig. 3D and fig. S9).
Li X., Mei Q., Yu Q., Wang M., He F., Xiao D., Liu H., Ge F., Yu X, & Li S. (2023). The TORC1 activates Rpd3L complex to deacetylate Ino80 and H2A.Z and repress autophagy. Science Advances, 9(10), eade8312.
Publication 2023
anti-IgG Antibodies Antibodies, Anti-Idiotypic Antibody Specificity Cell Extracts Goat Histone H2a Immunoblotting Immunoglobulins INO80 protein, human Lysine Mus Rabbits
2
Histone Octamer Reconstitution and rNCP Assembly
Bacterial expression vectors for histones H2A and H2B were purchased from Addgene (42,634 and 42,630, respectively). Plasmids to express X. laevis H3 (xH3) in pET-3d and xH4 in pET-3a were obtained from Professor Arrowsmith (University of Toronto). Introduction of C110A and K79C mutations in xH3 was performed by site-directed mutagenesis using QuikChange (Stratagene), and the plasmid was sequence-verified. Recombinant histones were purified from E. coli, and modified where indicated, before octamer assembly and subsequent refolding of rNCP with a 151 base pair 601 Widom DNA as previously described (Dyer et al, 2003 (link); Galloy et al, 2021 (link)). Briefly, the histones were purified from inclusion bodies under denaturing conditions on a 5-ml HiTrap SP FF (GE Healthcare) cation exchange column on a next-generation chromatography (NGC, Bio-Rad). Fractions containing the purified histone were pooled and dialyzed three times into 4 liters of water and 2 mM β-mercaptoethanol before lyophilization. The four histones were then unfolded into 20 mM Tris, pH 7.5, 7 M guanidine–HCl, and 10 mM DTT and mixed in equimolar ratios before octamer refolding into 2 M NaCl, 10 mM Tris, pH 7.5, 1 mM EDTA. Octamers were then purified on a Superdex 200 HiLoad 16/600 size exclusion column (GE Healthcare) and wrapped with the 151 base pair 601 Widom DNA to obtain rNCPs. Native gel analysis was used to validate the quality of the reconstitution.
Binda O., Kimenyi Ishimwe A.B., Galloy M., Jacquet K., Corpet A., Fradet-Turcotte A., Côté J, & Lomonte P. (2023). The TUDOR domain of SMN is an H3K79me1 histone mark reader. Life Science Alliance, 6(6), e202201752.
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Publication 2023
2-Mercaptoethanol Bacteria Base Pairing Chromatography Cloning Vectors Edetic Acid Escherichia coli Freeze Drying Galloy Guanidine Histone H2a Histones Inclusion Bodies Mutagenesis, Site-Directed Mutation Plasmids Sodium Chloride Tromethamine Xenopus laevis
3
Histone and FACT Complex Purification
Histones H2A, H2B, H3 and H4 were cloned into pET3a histone expression plasmid, overexpressed in E. coli, and purified from inclusion bodies50 (link). The purified histone ubH2B was purchased from KS-V Peptide Biological Technology Co., Ltd (Hefei, China), by total chemical synthesis utilizing standard solid phase peptide synthesis and peptide fragments ligation reactions to generate the desired protein51 (link). For the purification of recombinant FACT complex19 (link),27 (link), Sf9 cells (1.5–2 × 106/ml) were infected with baculovirus containing Flag-SPT16 and His6-SSRP1, and incubated at 27 °C for 72 h. The infected cells were collected and washed with ice-cold PBS, and then lysed in lysis buffer (150 mM NaCl, 20 mM Tris-HCl, pH 8.0, 5% glycerol, 1 mM PMSF). 10 mg (60 mL) of the cell extracts were incubated with 500 μL anti-Flag M2-agarose (Sigma, A2220) for 4 h at 4 °C, with the resin washed by lysis buffer. Bound proteins were eluted in the presence of 0.5 mg/ml Flag peptide (Sigma, F4799) and further purified by a Heparin HP column (GE Healthcare). The fractions containing FACT complex were dialyzed against BC-100 buffer (100 mM NaCl, 10 mM Tris-HCl, pH 8.0, 0.5 mM EDTA, 20% glycerol, 1 mM DTT, 1 mM PMSF) and stored at −80 °C.
Luo A., Kong J., Chen J., Xiao X., Lan J., Li X., Liu C., Wang P.Y., Li G., Li W, & Chen P. (2023). H2B ubiquitination recruits FACT to maintain a stable altered nucleosome state for transcriptional activation. Nature Communications, 14, 741.
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Publication 2023
Baculoviridae Biopharmaceuticals Buffers Cell Extracts Cells Common Cold Edetic Acid Escherichia coli FLAG peptide Glycerin Heparin Histone H2a Histones Ligation Peptide Fragments peptide V Plasmids Proteins Resins, Plant Sepharose Sf9 Cells Sodium Chloride SSRP1 protein, human Tromethamine
4
Protein Extraction and Western Blot Analysis
Cells were washed with ice-cold PBS, pelleted, and lysed in 50 mM Tris–HCl at pH 8.0, 150 mM NaCl, 0.5% SDS, and 1% NP-40 supplemented with a Halt Protease inhibitor cocktail (Thermo Fisher Scientific, Waltham, MA, USA) and Phosphatase inhibitor cocktails 2 and 3 (Sigma, St. Louis, MO, USA) on ice for 30 min. Lysates were then cleared by centrifugation for 10 min at 14,000× g. Total protein concentration was measured using a DC protein assay (Bio-Rad, Hercules, CA, USA) and 10–50 µg of protein was mixed with loading buffer. For the analysis of H2A and γH2AX, protein samples were separated on 4–15% Mini-PROTEAN® TGX™ Precast Protein Gels (Bio-Rad) and transferred to a PVDF membrane using Trans-Blot Turbo RTA Mini 0.2 µm PVDF Transfer Kit (Bio-Rad). The primary antibodies used were: Histone H2A Antibody II (#2578, Cell Signaling), Phospho-Histone H2A.X (Ser139) (20E3), and Rabbit mAb (#9718, Cell Signaling). HRP-conjugated anti-rabbit antibody (Sigma) was used as secondary antibody. Immuno-reactive bands were detected on the ChemiDoc MP system (Bio-Rad) using the Clarity Western ECL substrate (Bio-Rad).
Anisenko A., Nefedova A., Agapkina Y, & Gottikh M. (2023). Both ATM and DNA-PK Are the Main Regulators of HIV-1 Post-Integrational DNA Repair. International Journal of Molecular Sciences, 24(3), 2797.
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Publication 2023
Antibodies Antibodies, Anti-Idiotypic Biological Assay Buffers Cells Centrifugation Cold Temperature Gels H2AX protein, human Histone H2a Immunoglobulins Nonidet P-40 polyvinylidene fluoride Protease Inhibitors protein phosphatase inhibitor-2 Proteins Rabbits Sodium Chloride Tissue, Membrane Tromethamine
5
Immunofluorescent Staining of Paraffin Embedded Tissues
Tumor tissues that were to be used for immunofluorescent staining (IF) were fixed in formalin for one day at room temperature (RT). Tissues were then processed, dehydrated and embedded in paraffin using a tissue processor (RTPH-360, General Data Healthcare). IF was performed on 4 µm cut paraffin embedded tissues. Tissue sections were first deparaffinized and rehydrated. Antigen retrieval was then performed by boiling sections for 20 min in pH 9 antigen retrieval buffer (DAKO, S2368). Then, sections were permeabilized using 0.5 % triton X-100 (Sigma-Aldrich) in PBS (PBS-T) for 10 min at RT and then incubated in blocking buffer (3% BSA in PBS-T) for 30 min at RT. Sections were then incubated in blocking buffer containing the primary antibody (phosphorylated histone 2A (γH2AX) (Millipore, Amsterdam, The Netherlands, JBW301; 1:250)) for 90 min at RT. After washing, the sections were incubated in blocking buffer containing the secondary antibody for 45 min at RT. The sections were mounted using Vectashield containing DAPI (Vector Labs, Newark, CA, USA, H-1200).
Feijtel D., Reuvers T.G., van Tuyll-van Serooskerken C., de Ridder C.M., Stuurman D.C., de Blois E., Verkaik N.S., de Bruijn P., Koolen S.L., de Jong M, & Nonnekens J. (2023). In Vivo Efficacy Testing of Peptide Receptor Radionuclide Therapy Radiosensitization Using Olaparib. Cancers, 15(3), 915.
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Publication 2023
Antigens Buffers Cloning Vectors DAPI Fluorescent Antibody Technique Formalin Histone H2a Immunoglobulins Neoplasms Paraffin Embedding Tissues Triton X-100

Top products related to «Histone H2a»

1
Histone h2a by Cell Signaling Technology
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Histone H2A is a core histone protein that is a fundamental component of chromatin in eukaryotic cells. It plays a crucial role in the structural organization and regulation of DNA within the nucleus.
2
Ab18255 by Abcam
Sourced in United Kingdom
Ab18255 is a monoclonal antibody that targets a specific protein. It is designed for use in various laboratory applications, such as immunoassays and other experimental techniques. The product details and technical specifications are available on the Abcam website.
3
β actin by Merck Group
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β-actin is a protein that is found in all eukaryotic cells and is involved in the structure and function of the cytoskeleton. It is a key component of the actin filaments that make up the cytoskeleton and plays a critical role in cell motility, cell division, and other cellular processes.
4
Histone h2a by Abcam
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Histone H2A is a core histone protein that is a component of the nucleosome, the fundamental unit of chromatin. It plays a crucial role in the structural and functional organization of chromatin within the eukaryotic nucleus.
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Pvdf membrane by Merck Group
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Ab1791 by Abcam
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Ab1791 is a primary antibody targeting the protein Ki-67. It is suitable for use in immunohistochemistry and western blot applications. The product is provided as a liquid solution.
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Anti histone h2a antibody by Abcam
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Anti-Histone H2A antibody is a laboratory reagent used to detect and quantify the presence of Histone H2A protein in biological samples. It is a specific antibody that binds to the Histone H2A protein, allowing researchers to study its expression and localization in cells or tissues.
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Mesa green qpcr mastermix plus for sybr assay by Eurogentec
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MESA GREEN qPCR MasterMix Plus for SYBR Assay is a ready-to-use solution for quantitative real-time PCR (qPCR) analysis using SYBR Green detection. It contains all the necessary components, including a hot-start DNA polymerase, dNTPs, MgCl2, and SYBR Green I, to perform qPCR reactions.
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Anti histone h2a by Abcam
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Anti-Histone H2A is a laboratory reagent used to detect and quantify the presence of Histone H2A protein in biological samples. It is designed to specifically bind to Histone H2A, allowing for its identification and measurement.
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Histone h3 by Abcam
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Histone H3 is a core histone protein found in eukaryotic cells. It is involved in the structural organization of chromatin by forming nucleosomes, the fundamental units of chromatin.

More about "Histone H2a"

Histone H2A, a fundamental component of chromatin, plays a crucial role in regulating gene expression, DNA repair, and other essential cellular processes.
This core histone protein is responsible for the organization and compaction of DNA within eukaryotic cells, forming the nucleosome, the basic unit of chromatin.
Histone H2A, along with other histone proteins like Histone H3 and β-actin, contribute to the stability and structure of the nucleosome, directly influencing chromatin dynamics and biological functions.
Researchers studying Histone H2A often utilize techniques such as Western blotting with Anti-Histone H2A antibodies, qPCR with MESA GREEN qPCR MasterMix Plus for SYBR Assay, and other methods to investigate its role in gene regulation, DNA damage response, and epigenetic modifications.
Understanding the nuances of Histone H2A and its interactions with other chromatin-associated proteins is crucial for advancinig our knowledge of cellular processes.
PubCompare.ai's AI-driven platform offers a powerful tool to streamline and optimize Histone H2A research protocols, helping scientists effortlessly identify the best methodologies from literature, preprints, and patents.
Experience the future of scientific productivity and enhance your Histone H2A research with PubCompare.ai's innovative solutions.