product information Background ATP synthase is the universal enzyme that synthesizes ATP from ADP and phosphate using the energy stored in a transmembrane ion gradient. Immunogen KLH-conjugated synthetic peptide derived from available plant, algal (chloroplastic and mitochondrial) and bacterial sequences of beta subunits of F-type ATP synthases, including Arabidopsis thaliana chloroplastic ATP synthase subunit beta UniProt: P19366, TAIR: AtCg00480 and Arabidopsis thaliana mitochondrial ATP synthase subunit beta-1, UniProt: P83483, TAIR: At5g08670 as well as Chlamydomonas reinhardtii, UniProt: P06541 and A8IQU3 Host Chicken Clonality Polyclonal Clone Purity Total IgY Format Liquid at 23.3 µg/µl in PBS pH 8.0 + 0.02% sodium azide as preservative Quantity 100 µl Reconstitution Storage store at 4°C; make aliquots to avoid working with a stock. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from liquid material adhering to the cap or sides of the tubes. Tested applications Western blot (WB), immunolocalization (IL) Related products AS05 085 | anti-ATP synthase subunit beta, rabbit antibody AS03 030S | ATP synthase subunit beta protein standard for quantitation and positive control AS08 304 | anti-ATP synthase subunit alpha, rabbit antibody AS08 312 | anti-ATP synthase subunit gamma, rabbit antibody AS05 071 | anti-ATP synthase subunit c, rabbit antibodycollection of antibodies to proteins involved in bioenergetics of a plant cell Plant and algal protein extraction buffer Secondary antibodies Additional information The anti-AtpB antibody will detect the mitochondrial form of the F1 ATP synthase subcomplex, as well as the chloroplastic CF1 ATP synthase and most known bacterial F-type ATP synthases. Peptide used for antibody production is located in a beta sheet, which is partly exposed near the surface of the AtpB protein.

application information
Recommended dilution		1: 5 000 - 1: 8 000 (WB), 1: 500 for localization of native enzyme by immunogold (IL)
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		dicots including Glycine max, Vitis vinifera and monocots including Oryza sativa, Chlamydomonas reinhardtii, cyanobacteria, marine diatoms, Acinetobacter baumannii, Clostridium sp., bacteria including Yrsinia sp.
Not reactive in		archeal V-type ATP synthase
Additional information		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.

application information10 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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