V-ATPase, B | vacuolar H+-ATPase subunit B

AS14 2775 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Vigna radiata

Product Information

Immunogen

KLH-conjugated synthetic peptide chosen from Arabidopsis thaliana V-ATPase subunit B, isoform B1: UniProt: Q683E8, TAIR: AT1G76030, isoform B2 UniProt: Q9SZN1, TAIR: AT4G38510, isoform B3: UniProt: Q8W4E2 , TAIR: AT1G20260

Host Rabbit

Clonality Polyclonal

Purity Affinity purified serum in PBS, pH 7.4

Format Lyophilized

Quantity 50 µg

Reconstitution For reconstitution add 50 µl of sterile water.

Storage Store lyophilized/reconstituted at -20°C; 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 material adhering to the cap or sides of the tubes.

Tested applications Western blot (WB)

Recommended dilution 1 : 1000 (WB)

Expected | apparent MW

53 | 57 kDa (Vigna radiata)

Reactivity

Confirmed reactivity Arabidopsis halleri, Nicotiana tabaccum, Thellungiella salsuginea, Vigna radiata

Predicted reactivity

Acetabularia sp. , Arundo donax, Chlamydomonas reinhardtii, Cucumis sativus, Glycine soja, Gossypium mexicanum, Halostachys caspica, Haloxylon ammodendron, Hordeum vulgare, Medicago truncatula, Mesembryantheum crystallinum, Ostreococcus tauri, Oryza sativa, Panax ginseng, Physcomitrella patens, Pinus sylvestris, Populus trichocarpa, Pyrus sp., Ricinus communis, Theobroma cacao, Triticum aestivum, Zea mays, Zostera marina

Species of your interest not listed? Contact us

Not reactive in No confirmed exceptions from predicted reactivity are currently known.

Application examples

Application examples Applicaiton example

western blot using anti-V-ATPase subunit B antibodies

Proteins were separated by SDS-PAGE and transferred to an Immobilon-P membrane (Millipore) using Trans-Blot SD Semi-Dry Transfer Cell (Bio-Rad) with transfer buffer (100 mM Tris, 192 mM Glycine, 0.02% (w/v) SDS and 5% (v/v) methanol). After treatment with 1% blocking agent, the membrane filter was incubated with the primary antibody (1:1000) and then with horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (H+L) (Agrisera AS09 602, 1:25 000). Chemiluminescent reagent was used for detection of antigens. Chemiluminescence was detected with a Light-Capture II imaging device with a cooled CCD camera (Atto).

Samples:
1: 10 μg of 100,000 x g precipitate prepared from Arabidopsis thaliana 6 weeks old shoot.
2: 0.2 μg of vacuolar membrane enriched fraction prepared from Arabidopsis thaliana 6 weeks old shoot.
3: 2 μg of vacuolar membrane enriched fraction prepared from Arabidopsis thaliana 6 weeks old shoot.
4: 0.2 μg of vacuolar membrane enriched fraction prepared from Vigna radiata 4 days old hypocotyls. 5: 2 μg of vacuolar membrane enriched fraction prepared from Vigna radiata 4 days old hypocotyls.

Courtesy of Drs. Masayoshi Maeshima and Dr Shoji Segami, Nagoya University, Japan

Additional information

Antibodies will detect target protein in a few µg of a crude preparation loaded per well. If purified preparations of vacuolar and plasma membranes are used, one µg load per well should be sufficient.

Protein or membrane sample should be treated at 70°C for 10 min before loading on the gel.

Related products

collection of antibodies to vacuolar proteins

AS09 467 | Anti-V-ATPase subunit A | vacuolar H+-ATPase, rabbit antibodies

AS09 468 | Anti-V-ATPase subunit c | vacuolar H+-ATPase, subunit c (16 kDa), rabbit antibodies

AS09 497 | Anti- V-ATPase subunit D |V-type proton ATPase subunit D, rabbit antibodies

AS07 213 | Anti-V-ATPase subunit E of tonoplast H+ATPase, rabbit antibodies

AS09 499 | Anti-V-ATPase subunit H |V-type proton ATPase subunit H, rabbit antibodies

Plant protein extraction buffer

Background

V-ATPase, subunit B is a non-catalytic subunit of the peripheral V1 complex of vacuolar ATPase. Alternative names: Vacuolar proton pump subunit B1, vacuolar H+-ATPase subunit B, V-ATPase 57 kDa subunit.