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AtpB | beta subunit of ATP synthase (100 ĩl)

330 €

AS05 085  |  clonality: polyclonal  |  host: rabbit  |  reactivity: [global antibody] for plant, green alga, animal and bacterial F-type ATP synthases

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AS05 085

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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 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

antibody format  

rabbit;

polyclonal;

serum;

lyophilized

quantity  

100 µl

- for reconstitution add 100 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  

immunofluorescence (IF), Western blot (WB), Blue Native-PAGE  (BN-PAGE)

related products  

AS05 085-10 | anti-AtpB rabbit antibody, smaller pack size of AS05 085
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 antibody

AS08 312 | anti-ATP synthase subunit gamma antibody

AS05 071 | anti-ATP synthase subunit c antibody

recommended secondary antibody

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

application information
recommended dilution  

1:100 (IF), 1: 2000 - 1: 5 000 with standard ECL (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  

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  

dicots, including Vitis vinifera, and monocots:Triticum aestivum, algae, cyanobacteria, marine diatoms, Acinetobacter baumannii, Clostridium sp., bacteria including E.coli K-12, Trichodesmium erythraeum, Salmonella typhimurium, Yrsinia 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  

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


||| For applications or usage on species others than stated above Agrisera offers a payment-after-testing option. To learn more about this or for any questions on this product, please use the LiveChat option in the left menu bar or contact us at  support@agrisera.com

Do you have a matching secondary antibody?

AS09 602 | Goat anti-rabbit IgG (H&L), HRP conjugated

AS09 607 | Goat anti-rabbit IgG (H&L), ALP conjugated 

AS10 1489 | Rabbit anti-Chicken IgY (H&L), HRP conjugated 

AS09 606 | Goat anti-chicken IgY (H&L), ALP conjugated

For a complete list of Agrisera secondary antibodies


Detection reagents:

AS14 ECL-100 | AgriseraECL Bright 
AgriseraECL Bright for Western Blot detection is a high quality substrate for detection of horseradish peroxidase enzyme activity at a femtogram level. Its a ready to use 2 component system with low background and superior signal to nouse ratios.

AS14 TMB-HRP | AgriseraTMB HRP Peroxidase Microwell Substrate (100 ml) 
TMB based, one component, especially formulated with extreme sensitivity, HRP substrate for microwell application.