showForm Anti-AtpB (Beta subunits of ATP synthase) GLOBAL ANTIBODY

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AtpB | Beta subunit of ATP synthase (chloroplastic + mitochondrial) (rabbit)

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

AtpB | Beta subunit of ATP synthase (chloroplastic + mitochondrial) (rabbit) in the group Antibodies for Plant/Algal  / Global Antibodies at Agrisera AB (Antibodies for research) (AS05 085)

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Product Information

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 Rabbit
Clonality Polyclonal
Purity Serum
Format Lyophilized
Quantity 50 µl
Reconstitution For reconstitution add 50 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)
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)

Reactivity

Confirmed reactivity Arabidopsis thaliana, Bacillus cereus, Bryopsis corticulans, Chlamydomonas reinhardtii, Chlorella vulgaris, Chromochloris zofingiensis, Cyanidioschyzon merolae, Echinochloa crus/galli,  Escherichia coli, Helicobacter pylori, Hordeum vulgare, Glycine max, Lycopersicum esulentum, Moniliophthora perniciosa, Nannochloropsis salina, Neochloris oleoabundans (chlorophyta), Nicotiana bentamiana, Nicotiana tabacum, Oryza sp. (roots, leafs, pollen), Pheodactylum tricornutum CCAP 1055/1, Pisum sativum, Plasmodium berghei, Populus sp., Robinia pseudoacacia, Selaginella martensii, Spinacia oleracea, Toxoplasma gondii, Zea mays

Animal tissues from: cow, chicken, pig, rat, salmon, seal, Locusta migratoria
Predicted reactivity Acinetobacter baumannii, Algae, Cannabis sativa, 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

Application examples

Application examples 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 chemiluminescent substrate, using a Fuji LAS-3000 CCD (300s, standard sensitivity). 




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% blocking reagent  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% blocking solution for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with chemiluminescent 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

Additional information

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.

Anti-AtpB antibody was used as a loading control in Chlamydomonas reinhardtii and Synechocystis sp. PCC6803.

This product can be sold containing proclin if requested

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.

Related products

Related products
AS05 085-10 | Anti-AtpB rabbit antibody, smaller pack size of AS05 085, rabbitantibodies
AS05 085PRE | AtpB | beta subunit of ATP synthase, pre-immune serum
AS03 030 | Anti-AtpB hen antibody (developed to exactly the same peptide as rabbit antibody)
AS03 030S | ATP synthase subunit beta protein standard for quantitation and positive control
AS08 304 | Anti-ATP synthase subunit alpha, rabbit antibodies
AS08 312 | Anti-ATP synthase subunit gamma rabbit antibodies
AS05 071 | Anti-ATP synthase subunit c rabbit antibodies
AS16 3976 | Anti-AtpB | Beta subunit of ATP synthase, mitochondrial, rabbit antibodies

Background

Background

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

Product citations

Selected references Vojta and Fulgosi (2019). Topology of TROL protein in thylakoid membranes of Arabidopsis thaliana. Physiol Plant. 2019 Jan 20. doi: 10.1111/ppl.12927.
Roth et al. (2019). Regulation of Oxygenic Photosynthesis during Trophic Transitions in the Green Alga Chromochloris zofingiensis. Plant Cell. 2019 Feb 20. pii: tpc.00742.2018. doi: 10.1105/tpc.18.00742.
Aihara et al. (2019). Algal photoprotection is regulated by the E3 ligase CUL4-DDB1DET1. Nat Plants. 2019 Jan;5(1):34-40. doi: 10.1038/s41477-018-0332-5.
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.

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