Plant/Algal cell antibodies / Populus sp.

product information

ATP synthase is the universal enzyme that synthesizes ATP from ADP and phosphate using the energy stored in a transmembrane ion gradient.

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 AtCg00480  and Arabidopsis thaliana mitochondrial ATP synthase subunit beta-1 At5g08670 as well as Chlamydomonas reinhardtii P06541 and
A8IQU3

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.

Western blot (WB), immunolocalization (IL)

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 antibody

AS08 312 | anti-ATP synthase subunit gamma antibody

AS05 071 | anti-ATP synthase subunit c antibody

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

1: 5 000 - 1: 8 000 (WB), 1: 500 for localization of native enzyme by immunogold (IL)

53.9  kDa (Arabidopsis thaliana), 51.7 kDa (Synechocystis PCC 6803), 53.7 kDa (Spinacia oleracea)

Arabidopsis thalian, Spinacia oleracea, Synechocystis PCC 6803

dicots including Glycine max, Vitis vinifera and monocots including Hordeum vulgare, Oryza sativa, Chlamydomonas reinhardtii, cyanobacteria, marine diatoms, Clostridium sp., bacteria including Yrsinia sp.

archeal V-type ATP synthase

results of immunogold studies using anti-AtpB antibody are published in Andersson et al. (2009)

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 177:136-141

Johnson (2008) Altered expression of the chloroplast ATP synthase through site-directed mutagenesis in Chlamydomonas reinhardtii. Photosyn Res. 96(2):153-162.

application example

AtpB protein standard (AS03 030S) 0.03, 0.1, 0.26 pmol (1-3) and total protein from Trichodesmium IMS 101 extracted with PEB (AS08 300) were 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 (anti-hen IgY horseradish peroxidase conjugated, from Abcam) 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 the manufacturer's instructions. Images of the blots were obtained using a CCD imager nd Quantity One software  Exposure time was 10 seconds.
Note: Optimal quantitation is achieved using moderate sample loads per gel lane, generally 0.5 to 2.5 ug total protein, depending on the abundance of the target protein.