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

411 €

AS09 503 | clonality: polyclonal | host: rabbit | reactivity: Acetabularia sp. Hordeum vulgare, Oryza sativa, Pinus sylvestris, Pyrus sp. ,Vigna radiata

PRODUCT INFORMATION IN PDF

product information

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.

immunogen

purified native V-ATPase, subunit B from Vigna radiata

antibody format

rabbit

polyclonal,

ammonium sulfate purified IgG,

quantity

100 µl

storage

store 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

ELISA (ELISA), western blot (WB)

related products

collection of antibodies to vacuolar proteins

AS09 467 | anti-V-ATPase subunit A | vacuolar H+-ATPase

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

ASS09 497 | anti- V-ATPase subunit D |V-type proton ATPase subunit D

AS07 213 | anti-V-ATPase subunit E of tonoplast H+ATPase

AS09 499 | anti-V-ATPase subunit H |V-type proton ATPase subunit H

additional information

0.1 % sodium azide is added as preservative. For antibody re-suspending information check the tube lable.

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.

Protocol for isolation of vacuolar membranes can be found here.

application information
recommended dilution		1: 8000 (ELISA), 1: 2000 with standard ECL (WB)
expected \| apparent MW		53 \| 57 kDa (Vigna radiata)
confirmed reactivity		Acetabularia sp. Hordeum vulgare, Oryza sativa, Pinus sylvestris, Pyrus sp. ,Vigna radiata
predicted reactivity		dicots including: Arabidopsis thaliana, Gossypium mexicanum, monocots including: Hordeum vulgare, Oryza sativa, trees: Populus trichocarpa, moss: Physcomitrella patens, algae: Chlamydomonas reinhardtii, microalgae:Ostreococcus tauri
not reactive in		no confirmed exceptions from predicted reactivity known in the moment
additional information		Protein or membrane sample should be treated at 70°C for 10 min before loading on the gel. Diluted antibody solution can be used 2 to 3 times within one month if it contains 0.1 % sodium azide as preservative and is stored at -20ºC to -80ºC. Manufactured by Operon Biotechnologies.
selected references		Kawamura et al. (2000). Tissue specificity of E subunit isoforms of plant vacuolar H(+)-ATPase and existence of isotype enzymes. J.Biol. Chem. 275:6515-6522. Nakanishi & Maeshima (1998). Molecular cloning of vacuolar H(+)-pyrophosphatase and its developmental expression in growing hypocotyl of mung bean. Plant Physiol. 116:589-597. Smart et al. (1998). Genes involved in osmoregulation during turgor-driven cell expansion of developing cotton fibers are differentially regulated. Plant Physiol. 116:1539-1549. Matsuura-Eno et al. (1992). Mechanism of the Decline in Vacuolar H -ATPase Activity in Mung Bean Hypocotyls during Chilling. Plant Physiology 100:718-722.

application example

65 µg/lane of purified vacuolar membranes from Vigna radiata L. (1,3) and purified V-ATPase, 7.4 µg/lane (2,4) were separated on 12 % SDS-PAGE and blotted 1h to PVDF membrane (40 min. at 10 V using BioRad semidry transfer). Filters were blocked 1h with 5 % low-fat milk powder in TBS-T (0.05% Triton X.100). Membranes were washed 5 times with TBS-T, each time in a fresh polystyrene box and probed with anti-V-ATPase subunit B antibodies (AS09 503, 1:2000, 1h) and secondary anti-rabbit (1:2000, 1 h). All steps were performed in RT with agitation.

CBB - staining with Coomasie blue - left panel. Immunoblot - western blot detectoin - right panel.

western blot detection using anti-V-ATPase subunit B antibodies

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