AtpB | Beta subunit of ATP synthase, (chloroplastic + mitochondrial) (rabbit antibody)
AS05 085 | Clonality: Polyclonal | Host: Rabbit | Reactivity: [global antibody] for plant, green alga, animal and bacterial F-type ATP synthases
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|Recommended dilution||1 : 100 (IF), 1 : 5000 (BN-PAGE), 1 : 2000-1 : 5 000 (WB)|
|Expected | apparent MW||
53.9 kDa (Arabidopsis thaliana), 51.7 kDa (Synechocystis PCC 6803), 53.7 kDa (Spinacia oleracea)
|Confirmed reactivity||Arabidopsis thaliana, Bacillus cereus, Chlamydomonas reinhardtii, Chlorella vulgaris, Cyanidioschyzon merolae, Escherichia coli, Helicobacter pylori, Hordeum vulgare, Glycine max, Lycopersicum esulentum, Moniliophthora perniciosa, Neochloris oleoabundans (chlorophyta), Nicotiana bentamiana, Nicotiana tabacum, Oryza sp. (roots, leafs, pollen), Pheodactylum tricornutum CCAP 1055/1, Plasmodium berghei, Populus sp., Selaginella martensii, Spinacia oleracea, Zea mays
Animal tissues from: cow, chicken, pig, rat, salmon, seal, Locusta migratoria
|Predicted reactivity||Acinetobacter baumannii, Algae, Clostridium sp., Cyanobacteria, E.coli K-12, Nicotiana plumbaginifolia, Saccharomyces cerevisiae, Salmonella typhimurium, Trichodesmium erythraeum, Triticum aestivum, Vitis vinifera, Zosteria marina, Yrsinia sp.|
|Not reactive in||Archeal V-type ATP synthase|
Blue Native gel electrophoresis (BN-PAGE) has been performed on samples solubilized with digitonin (4:1) and loaded at 100 µg/well. Gel thickness was 2 mm with 4.5-16 % gradient.
Antibody is recognizing mitochondrial form of AtpB Subota el. al (2011).
This antibody can be used as a loading control for bacteria, Bacillus cereus.
|Selected references||Fu et al. (2017). Intracellular spectral recompositioning of light enhances algal photosynthetic efficiency. Sci Adv. 2017 Sep 1;3(9):e1603096. doi: 10.1126/sciadv.1603096. (loading control diatoms)
Shin et al. (2017), Complementation of a mutation in CpSRP43 causing partial truncation of light-harvesting chlorophyll antenna in Chlorella vulgaris. Sci Rep. 2017 Dec 20;7(1):17929. doi10.1038/s41598-017-18221-0.
Wang and Auwerx (2017). Systems Phytohormone Responses to Mitochondrial Proteotoxic Stress. Mol Cell. 2017 Nov 2;68(3):540-551.e5. doi: 10.1016/j.molcel.2017.10.006.
Cantrell and Peers (2017). A mutant of Chlamydomonas without LHCSR maintains high rates of photosynthesis, but has reduced cell division rates in sinusoidal light conditions. PLoS One. 2017 Jun 23;12(6):e0179395. doi: 10.1371/journal.pone.0179395.
Nikolova et al. (2017). Temperature induced remodeling of the photosynthetic machinery tunes photosynthesis in a thermophyllic alga. Plant Physiol. 2017 Mar 7. pii: pp.00110.2017. doi: 10.1104/pp.17.00110.
Jiang et al. (2017). IFT57 stabilizes the assembled intraflagellar transport complex and mediates transport of motility-related flagellar cargo. J Cell Sci. 2017 Mar 1;130(5):879-891. doi: 10.1242/jcs.199117.
Li et al. (2016). Characterization of a novel β-barrel protein (AtOM47) from the mitochondrial outer membrane of Arabidopsis thaliana. J Exp Bot. 2016 Nov;67(21):6061-6075. Epub 2016 Oct 6.
Jallet et al. (2016). Photosynthetic physiology and biomass partitioning in the model diatom Phaeodactylum tricornutum grown in a sinusoidal light regime. Algal research, doi:10.1016/j.algal.2016.05.014.
Ferroni et al. (2016). Light acclimation in the lycophyte Selaginella martensii depends on changes in the amount of photosystems and on the flexibility of the light-harvesting complex II antenna association with both photosystems. New Phytol. 2016 Apr 5. doi: 10.1111/nph.13939.
Heinnickel et al. (2016). Tetratricopeptide repeat protein protects photosystem I from oxidative disruption during assembly. Proc Natl Acad Sci U S A. 2016 Mar 8;113(10):2774-9. doi: 10.1073/pnas.1524040113
2 µg of total protein extracted with PEB (AS08 300) from leaf tissue of (1) Arabidopsis thaliana, (2) Spinacia oleracea, (3) Lycopersicon esculentum, (4) Glycine max, (5) Populus sp., (6) Zea mays and (7) Hordeum vulgare were separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to nitrocellulose. In parallel a dilution row (a-g: 10 - 5 - 2.5 - 1.25 - 0.63 - 0.32 - 0.16 µg protein/lane) from sample 1 (Arabidopsis) was processed. Filters were blocked 1h with 2% low-fat milk powder in TBS-T (0.1% TWEEN 20) and probed with anti-AtpB (AS08 085, 1:5000, 1h) and secondary anti-rabbit (1:10000, 1 h) antibody (HRP conjugated, recommended secondary antibody AS09 602) in TBS-T containing 2% low fat milk powder. Antibody incubations were followed by washings in TBS-T (15, +5, +5, +5 min). All steps were performed at RT with agitation. Signal was detected with standard ECL (Invitrogen) using a Fuji LAS-3000 CCD (300s, standard sensitivity).
Application example 2
2 µg of total protein from (1) cow muscle, (2) chicken muscle, (3) pig muscle, (4) rat liver, (5) salmon muscle, (6) seal muscle, (8) Arabidopsis thaliana, (9) Zea mays extracted with Protein Extration Buffer, PEB (AS08 300) and separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to PVDF. Blots were blocked immediately following transfer in 2% ECL Advance blocking reagent (GE Healthcare) in 20 mM Tris, 137 mM sodium chloride pH 7.6 with 0.1% (v/v) Tween-20 (TBS-T) for 1h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1: 50 000 for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (Agrisera anti-rabbit IgG horse radish peroxidase conjugated, AS09 602) diluted to 1:50 000 in 2% ECL Advance blocking solution for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with ECL Advance detection reagent according to the manufacturers instructions. Images of the blots were obtained using a CCD imager (FluorSMax, Bio-Rad) and Quantity One software (Bio-Rad). Exposure time was 30 seconds.
M - molecular weight marker
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