AtpB | Beta subunit of ATP synthase (chloroplastic + mitochondrial) (chicken antibody)
AS03 030 | Clonality: Polyclonal | Host: Chicken | Reactivity: [global antibody] for plant and bacterial F-type ATP synthases
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|Recommended dilution||1 : 500 for localization of native enzyme by immunogold (IL), 1 : 5 000-1 : 8 000 (WB)|
|Expected | apparent MW||
53.9 kDa (Arabidopsis thaliana), 51.7 kDa (Synechocystis PCC 6803), 53.7 kDa (Spinacia oleracea)
|Confirmed reactivity||Arabidopsis thaliana, Hordeum vulgare, Spartina alterniflora, Spinacia oleracea, Synechocystis PCC 6803, Synechococcus PCC 7942, beef muscle, Rat liver|
|Predicted reactivity||Acinetobacter baumannii, Chlamydomonas reinhardtii, , Clostridium sp., Cyanobacteria, Glycine max, Oryza sativa, marine Diatoms, Vitis vinifera, Yersinia sp.|
|Not reactive in||Archeal V-type ATP synthase|
Results of immunogold studies using anti-AtpB antibody are published in Andersson et al. (2009).
|Selected references||Quesada et al. (2011). Arabidopsis RUGOSA2 encodes an mTERF family member required for mitochondrion, chloroplast and leaf development. Plant J. Nov;68(4):738-53. doi: 10.1111/j.1365-313X.2011.04726.x. Epub 2011 Sep 13.
Andersson et. al (2009). Co-localization of P-glycerate kinase, P-ribulokinase, ADP-glucose pyrophosphorylase and Rubisco activase with CF1 in pea leaf chloroplasts. Plant Science 177:136-14. (immunolocalization)
Morash et al. (2007). Macromolecular dynamics of the photosynthetic system over a seasonal developmental progression in Spartina alterniflora. Canadian J. of Botany, 2007, 85(5): 476-483, 10.1139/B07-043.
10 µg of total protein from samples such as beef muscle (1), rat liver (2), Arabidopsis thaliana leaf (3), Hordeum vulgare leaf (4), Synechocystis PCC 6803 total cell (5), Synechococcus PCC 7942 total cell (6) were extracted with Protein Extraction Buffer PEB (AS08 300). Samples were diluted with 1X sample buffer (NuPAGE LDS sample buffer (Invitrogen) supplemented with 50 mM DTT and heat at 70C for 5 min and keep on ice before loading. Protein samples were separated on Bolt 4-12% Tris-Bis gel (Invitrogen) LDS-PAGE and blotted for 1h to 1.5h on PVDF using Bolt Mini Blot Module. Blot was blocked immediately following transfer in 2% blocking reagent (GE RPN 2125; Healthcare) dissolved 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. Blot was incubated in the primary antibody at a dilution of 1: 5 000 (in blocking reagent) for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, and then washed 1x15 min and 3x5 min with TBS-T at room temperature with agitation. Blot was incubated in secondary antibody (anti-chicken IgY horse radish peroxidase conjugated) diluted to 1:20 000 in blocking reagent for 1h at room temperature with agitation. The blot was washed as above. Signals in the blot were detected using Lumigen ECL Ultra Reagent (Lumigen TMA-6, Lumigen), and visualized using the Molecular Imager VersaDoc MP 4000 System (Bio-Rad) and Quantity One software (Bio-Rad). Exposure time was 30 seconds.
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