H1 | Histone H1
AS11 1801 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, N. tabacum

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Product Information
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 µl 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)
Recommended dilution
1 : 100-1 : 500 (ICC), 1 : 5000 (WB)
Expected | apparent MW
15 | 17 kDa
Reactivity
Confirmed reactivity
Arabidopsis thaliana, Nicotiana tabacum, Triticum aestivum
Predicted reactivity
Lathyrus sativus, Phaseolus vulgaris, Pisum sativum, Solanum lycopersicum, Vicia faba
Species of your interest not listed? Contact us
Species of your interest not listed? Contact us
Not reactive in
No confirmed exceptions from predicted reactivity are currently known.
Application examples
Application examples
Western blot

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

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.

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

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