H3 | Histone H3 (chicken antibody)
AS15 2855 | Clonality: Polyclonal | Host: Chicken | Reactivity: A. thaliana, C. reinhardtii, L. esculentum, T. aestivum | cellular [compartment marker] of nucleoplasm
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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)
15 | 17 kDa
Arabidopsis thaliana, Brassica napus, Brassica oleracea, Capsicum annuum, Chlamydomonas acidophila, Hordeum vulgare, Medicago sativa, Nannochloropsis gaditana, Nicotiana tabacum, Oryza sativa, Physcomitrium patens, Pinus pinaster, Pisum sativum, Salicornia europaea, Solanum lycopersicum, Solanum sogarandinum, Solanum tuberosum, Triticum aestivum, Zea mays, Vicia faba, Vitis vinifera, Volvox sp.
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From left to right: 2 wells of nuclear + cytoplasmic enriched protein, 2 wells of total proteins of Chlamydomonas reinhardtii protein saturated in 8M urea were separated on 15% SDS-PAGE and blotted for 1hour to 0.2 µm nitrocellulose at 100V using wet transfer system. Blots were blocked with 0.5% cold fish gelatin for 1hr at room temp with agitation. Blot was incubated in the primary antibody (anti-H3) at a dilution of 1:2500 for an hour at RT with agitation. The blots were washed with 3X 15min TBS-TT at RT with agitation. Blots as incubated in the secondary antibody (AS11 1827, Goat anti-Chicken IgY (H&L), DyLight® 800 conjugated, Agrisera) 1:5000 dilution for 30min at RT with agitation and washed 1X with TBSTT for 15min, 1X with TBST for 15min before scanning with the ODyssey IRD scanner.
Courtesy of Dr. Betty Chung, University of Cambridge, United Kingdom
Reactant: Chlamydomonas reinhardtii (Green Alga)
Application: Western Blotting
Pudmed ID: 31366981
Journal: Sci Rep
Figure Number: 2C
Published Date: 2019-07-31
First Author: Chung, B. Y. W., Valli, A., et al.
Impact Factor: 4.13Open Publication
Expression and differential localisation of AGO2 and AGO3. (a) Antigen sites utilised for peptide-based antibody production. (b) Western blots for total and ?-2/3-PAZ-immunoprecipitated product using extracts from either the parental (P) line or ago3-25 (M). At-AGO1 antibody was utilized as an IP negative control. (c) Differential localisation of Cr-AGO2 and 3. Total protein isolated from cytoplasmic, total and nuclear fractions are indicated as C, T and N, respectively. Extracts from either Parental or ago3-25 strain are labelled P and M, respectively. Protein bands corresponding to AGO2/3 or Histone 3 (H3) were indicated.
Histone 3 (H3) located in nuclei, incorporated into chromatin. Present in nucleosome together with H2A, H2B and H4.
Chung et al. (2019) Distinct roles of Argonaute in the green alga Chlamydomonas reveal evolutionary conserved mode of miRNA-mediated gene expression. Sci Rep. 2019 Jul 31;9(1):11091. doi: 10.1038/s41598-019-47415-x.
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