H1 | Histone H1

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AS11 1801  |  Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity: A. thaliana, N. tabacum


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Item No:
AS11 1801

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product information

Histone 1 (H1) is a protein located in nuclei, incorporated into chromatin. 

Immunogen native H1 protein purified from Nicotiana tabaccum (H1A, H1B H1C,D,E,F)
Host Rabbit
Clonality Polyclonal
Purity Affinity purified serum in PBS, pH 7.4
Format Lyophilized
Quantity 50 µg
Reconstitution For reconstitution add 50 ĩl of sterile water.
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 Chromatin Immunoprecipitation (IP) (ChIP), Western blot (WB), Immunocytochemistry (ICC) (ICC)
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collection of antibodies to DNA/RNA/cell cycle

Plant protein extraction buffer

Secondary antibodies

Additional information
application information
Recommended dilution 1 : 100-1 : 500 (ICC), 1 : 5000 (WB)
Expected | apparent MW

15 | 17 kDa

Confirmed reactivity Arabidopsis thaliana, Nicotiana tabacum, Triticum aestivum
Predicted reactivity Lathyrus sativus, Phaseolus vulgaris, Pisum sativum, Solanum lycopersicum, Vicia faba
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.

Protocol for immunostatining in whole-mount plant tissues: An efficient method for quantitative, single-cell analysis of chromatin modification and nuclear architecture in whole-mount ovules in Arabidopsis Wenjing She, Daniel Grimanelli, Célia Baroux Journal of Vizualized Experiments (JoVE), in press

Selected references Wollmann et al. (2017). The histone H3 variant H3.3 regulates gene body DNA methylation in Arabidopsis thaliana. Genome Biol. 2017 May 18;18(1):94. doi: 10.1186/s13059-017-1221-3.
She and Baroux (2015). Chromatin dynamics in Pollen Mother Cells underpin a common scenario at the somatic-to-reproductive fate transition of both the male and female lineages in Arabidopsis. Front. Plant Sci. | doi: 10.3389/fpls.2015.00294.
She et al. (2013). Chromatin reprogramming during the somatic to-reproductive cell fate transition in plants. Development Oct;140(19):4008-19. doi: 10.1242/dev.095034. Epub 2013 Sep 4. (Arabidopsis thaliana, immunostaining)

application example

western blot

western blot using plant anti-H1 antibodies

50 µl of a total protein from Arabidopsis thaliana leafs (wt and single, double and triple H1 mutants as well as overexpressed H1 as a GFP fusion) extracted in a following way: samples were grinded in LN2, suspended in 1xSDS buffer (sample:buffer 1:5), sonicated (10 min., max. power, in ice-cooled sonicating bath (BioRuptor, Diagenode, Belgium) and were separated on 15 % SDS-PAGE and blotted 2h to PVDF(Millipore Westran). Blots were blocked with 5 % non-fat milk TBST for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 5 000 for  over night at 4°C with agitation. The antibody solution was decanted and the blot was rinsed  and washed four times for 10 min in TBS-T at RT with agitation in 2.5 % non-fat milk in YBST. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated,from Agrisera, AS09 602) diluted to 1:10 000 in for 1h at RT with agitation. The blot was washed as above and developed for 5 min with a home made ECL. Exposure time was 5 min.

Courtesy of Dr. Maciej Kotliński, Institute of Biochemistry and Biophysics of Polish Academy of Sciences in Warsaw, Poland

immunolocalization in plant tissue using anti-H1 antibodies
Whole-mount immunostaining on Arabidopsis thaliana ovule primordia stage 2-II. Steps involved: clarification (methanol/xylene), cell wall digestion and permeabilization before application of the primary, then secondary antibody for 12-14 hours at 4°C; fixation: BVO buffer: buffer from (Bauwens and Van Oostveld 1996), 2mM EGTA pH7.5, 10% DMSO, 1% Tween in PBS (containg 1% formaldehyde) for 30 min. rotating/shaking plate RT; blocking: none; counterstaining: propidium iodide and mounted in Prolong Gold (Invitrogen); primary antibody dilution: 1:200 in PBS + 0.2% Tween-20; secondary antibody dilution: 1:200 at 4°C, 24h, goat anti-rabbit IgG Alexa 488 conjugated (Molecular Probes (A11008)). 

H1 immunostaining in Arabidopsis thaliana ovule primordia stage 2-II is in accordance with H1.1-GFP expression pattern (She et al 2013 et al. 2013).

Courtesy of Dr. Kinga Rutowicz, IBB PAS, Warsaw, Poland, with the technical assistance of Drs. Wenjing She and Célia Baroux, University of Zürich, Switzerland.

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