AtpB | Beta subunit of ATP synthase, chloroplastic + mitochondrial (rabbit antibody) (10 µl)

AS05 085-10 | Clonality: Polyclonal | Host: Rabbit | Reactivity: [global antibody] for plant, green alga and bacterial F-type ATP synthases

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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 AtCg00480 and Arabidopsis thaliana mitochondrial ATP synthase subunit beta-1 At5g08670 as well as Chlamydomonas reinhardtii P06541 and A8IQU3
Host		Rabbit
Clonality		Polyclonal
Clone
Purity		Serum
Format		Lyophilized
Quantity		10 µl
Reconstitution		For reconstitution add 10 µl of sterile water.
Storage		Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.
Tested applications		Blue Native-PAGE (BN-PAGE), Immunofluorescence (IF), Western blot (WB)
Related products		AS05 085 \| anti-AtpB \| beta subunit of ATP synthase (100 µl), rabbit antibody AS03 030 \| anti-AtpB (developed to exactly the same peptide as rabbit antibody), chicken 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 AS16 3976 \| anti-AtpB \| Beta subunit of ATP synthase, mitochondrial, rabbit antibody Plant and algal protein extraction buffer Secondary antibodies
Additional information		This product can be sold containing proClin if requested. 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 : 100 (IF), 1 : 2000-1 : 5 000 (WB) 1 : 5000 (BN-PAGE)
Expected \| apparent MW		53.9 kDa (Arabidopsis thaliana), 51.7 kDa (Synechocystis PCC 6803), 53.7 kDa (Spinacia oleracea)
Confirmed reactivity		Agostis stolonifera cv. ‘Penncross’, Arabidopsis thaliana, Bacillus cereus, Chlamydomonas reinhardtii, Hordeum vulgare, Glycine max, Lycopersicum esulentum, Oryza sp. (roots, leafs, pollen), Nicotiana bentamiana, Nicotiana tabacum, Populus sp., Spinacia oleracea, Zea mays. Animal tissues from: cow, chicken, pig, rat, salmon, seal, Locusta migratoria
Predicted reactivity		Acinetobacter baumannii, Algae, Cannabis sativa, Clostridium sp., Diatoms, Cyanobacteria, E.coli K-12, Vitis vinifera, Yersinia sp.
Not reactive in		Archeal V-type ATP synthase
Additional information		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		Jespersen et al. (2017). Metabolic Effects of Acibenzolar-S-Methyl for Improving Heat or Drought Stress in Creeping Bentgrass. Front Plant Sci. 2017 Jul 11;8:1224. doi: 10.3389/fpls.2017.01224. eCollection 2017. (western blot, Agostis stolonifera cv. ‘Penncross’) Rurek et al. (2015). Biogenesis of mitochondria in cauliflower (Brassica oleracea var. botrytis) curds subjected to temperature stress and recovery involves regulation of the complexome, respiratory chain activity, organellar translation and ultrastructure. Biochim Biophys Acta. 2015 Jan 21. pii: S0005-2728(15)00016-X. doi: 10.1016/j.bbabio.2015.01.005. Eom et al. (2014). Bacillus subtilis HJ18-4 from Traditional Fermented Soybean Food Inhibits Bacillus cereus Growth and Toxin-Related Genes. J Food Sci. 2014 Nov;79(11):M2279-87. doi: 10.1111/1750-3841.12569. Epub 2014 Oct 30. Lintala et al. (2013). Arabidopsis tic62 trol mutant lacking thylakoid bound ferredoxin-NADP+ oxidoreductase shows distinct metabolic phenotype. Mol Plant Sep 16. Teng et al. (2013). Mitochondrial Genes of Dinoflagellates Are Transcribed by a Nuclear-Encoded Single-Subunit RNA Polymerase. PLOS ONE, June 2013. (immuofluorescence) Rasala et al. (2013). Expanding the spectral palette of fluorescent proteins for the green microalga Chlamydomonas reinhardtii. Plant J. March 23. Heinnickel et al. (2013). Novel thylakoid membrane greencut protein cpld38 impacts accumulation of the cytochrome b6f complex and associated regulatory processes. J. Biol. Chem. Jan 9.

application example

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

western blot detection of AtpB in animal and plant tissue

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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Accessories: AtpB | Beta subunit of ATP synthase, chloroplastic + mitochondrial (rabbit antibody) (10 µl)

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