H3 | Histone H3 (100 ĩl)

AS10 710-100 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, S. lycopersicum, V. faba, P. patens, S. europaea, Z. mays, Ch. reinhardtii | cellular [compartment marker] of nucleoplasm

H3 | Histone H3 (100 ĩl) in the group Plant/Algal Antibodies / DNA/RNA/Cell Cycle / Nuclear signaling at Agrisera AB (Antibodies for research) (AS10 710-100)


407 €

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product information

Histone 3 (H3) located in nuclei, incorporated into chromatin. Present in nucleosome together with H2A, H2B and H4.


KLH-conjugated synthetic peptide derived from known H3 sequences, inluding Arabidopsis thaliana H3.3 P59169 (At4g40030,At4g40040,At5g10980), H3.2 P59226(At1g09200,At3g27360,At5g10390,At5g10400,At5g65360), H3-like 2 Q9FXI7 (At1g19890)

Host Rabbit
Clonality Polyclonal
Purity Serum
Format Lyophilized
Quantity 2 x 50 ĩl
Reconstitution For reconstitution add 50 ĩl of sterile water to each tube.
Storage Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.
Tested applications Western blot (WB), Immunocytochemistry (ICC) (ICC)
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AS10 710PRE | H3 | histone H3, pre-immune serum

collection of antibodies to DNA/RNA/cell cycle

Plant and algal protein extraction buffer

Secondary antibodies

Additional information Cellular [compartment marker] of nucleoplasm, loading control antibody for Chlamydomonas reinhardtii
application information
Recommended dilution 1 : 100-1 : 500 (ICC), 1 : 5000 (WB)
Expected | apparent MW

15 | 17 kDa

Confirmed reactivity Arabidopsis thaliana, Brassica oleracea, Capsicum annuum, Chlamydomonas acidophila, Chlamydomonas reinhardtii, Physcomitrella patens, Salicornia europaea, Solanum lycopersicum, Solanum sogarandinum, Solanum tuberosum, Vicia faba, Zea mays
Predicted reactivity Brassica napus, Hordeum vulgare, Nicotiana tabacum, Medicago sativa, Oryza sativa, Pisum sativum, Pinus pinaster, Triticum aestivum, Zea mays, Vitis vinifera, Volvox sp.
Not reactive in No confirmed exceptions from predicted reactivity are currently known.
Additional information

Protocol for isolation of cytosolic and nuclear fractions can be found here.

Specific fluorescence in ICC has been observed for interphase nuclei as well as around centromer region (where Ser10 of histone H3 is phosphorylated) in mitotic chromosomes.

Selected references Spijkerman et al. (2014). CO2 acquisition in Chlamydomonas acidophila is influenced mainly by CO2, not phosphorus, availability. Photosynth Res. 2014 Sep;121(2-3):213-21. doi: 10.1007/s11120-014-0016-6. Epub 2014 Jun 7.
Rihan et al. (2014). The effect of molybdenum on the molecular control of cold tolerance in cauliflower (Brassica oleracea var. botrytis) artificial seeds. Plant Cell, Tissue and Organ Culture (PCTOC) April 2014
Szabala et al. (2014). Accumulation of acidic SK3 dehydrins in phloem cells of cold- and drought-stressed plants of the Solanaceae. Planta, Jan 7.
Wobbe and Nixon (2013). The mTERF protein MOC1 terminates mitochondrial DNA transcription in the unicellular green alga Chlamydomonas reinhardtii. Nucleic Acids Res. May 6.

application example


western blot using anti-plant H3, histone 3 antibodies

1.2 μg of Arabidopsis thaliana chromatin-enriched fraction (1) and 3.75 µg of total protein from 4-weeks-old Arabidopsis thaliana leaves (2), and were separated on 12% SDS-PAGE and blotted 50 mins to Immobilon-P (Millipore, semi-dry) PVDF membrane. Blots were blocked immediately following transfer in MTBS-T (5% milk) for 30 mins at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1:5000 for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed 3 times for 3 min in TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (anti- IgG horse radish peroxidase conjugated, from Agrisera, AS09 602) diluted to 1:20 000 for 30 mins at room temperature with agitation. The blots were washed as above and developed for 5 min with ECL detection reagent (Roche) according to the manufacturers instructions. Exposure time was 30 seconds. Double band in chromatine-enriched fraction (1) has been outcompeted in peptide neutralization assay by peptide used to elicit H3 antibodies. Chromatin izolation was carried out as described (Zilberman et al. 2008) with minor modifications.

Courtesy of Weronika Sura and Dr. Piotr A. Ziolkowski (Department of Biotechnology, Adam Mickiewicz University, Poznan, Poland)



western blot on Chlamydomonas using anti-H3 antibody

30 μg of Chlamydomonas reinhardtii total (“Cells”) or nuclei-enriched (“Nuclei”) proteins were separated on a 13% SDS-PAGE gel and blotted 90 minutes to AmershamTM HybondTM-ECL (pore size 0.2 μm). Membranes were blocked in TBST 5% milk for one hour at room temperature and incubated at 4°C overnight in TBST 5% milk with the Agrisera
primary H3 antibody at a dilution of 1:5000. After three washes in TBST, blots were incubated one hour at room temperature with Agrisera HRP-conjugated Goat anti-rabbit IgG (H&L) diluted 1:50000 in TBST 5% milk.

Signal was visualized with standard ECL on Amersham HyperfilmTM ECL after 1 minute exposure. Molecular weight is determined by the Broad Range Protein Molecular Weight Marker (Promega).

Courtesy of Dr. Leonardo Magneschi, Hippler’s Lab, Institut für Biologie und Biotechnologie der Pflanzen (IBBP), Westfälische Wilhelms-Universität Münster, Germany


ICC using anti plant histone 3 antibody

Immunocytochemical assays were performed according to the method described earlier (Rybaczek and Maszewski 2006). Excised apical parts of Vicia faba roots (1.5 mm long) were fixed for 45 min (18°C) in PBS-buffered 3.7% paraformaldehyde, washed several times with PBS and placed in a citric acid-buffered digestion solution (pH 5.0; 37°C for 45 min) containing 2.5% pectinase (Fluka), 2.5% cellulase (Onozuka R-10; Serva) and 2.5% pectoliase (ICN). After removing the digestion solution, root tips were washed 3 times in PBS, rinsed with distilled water and squashed onto Super Frost Plus glass slides (Menzel-Gläser). Air-dried slides were pretreated with PBS-buffered 5% BSA at 20°C for 50 min and incubated overnight in a humidified atmosphere (4°C) with rabbit antibody raised against H3 histone (Agrisera), dissolved in PBS containing 1% BSA (at a dilution of 1:50). Following incubation, slides were washed 3 times with PBS and incubated for 1 h (18°C) with Agrisera secondary goat anti-rabbit IgG DyLight®488 antibody (AS09 633, 1:1000). Nuclear DNA was stained with 4’,6-diamidino-2-phenyl-indole (DAPI, 0.4 μg/ml; Sigma-Aldrich). Following washing with PBS, slides were air dried and embedded in Vectashield Mounting Media for Fluorescence (Vector Laboratories). Observations were made using Optiphot-2 fluorescence microscope (Nikon) equipped with B-2A filter (blue light; λ ≈ 495 nm) for DyLight-conjugated antibodies and UV-2A filter (UV light; λ ≈ 365 nm) for DAPI. All images were recorded at exactly the same time of integration using DXM 1200 CCD camera.

Courtesy Dr. Dorota Rybaczek, Lodz University, Poland

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