Plant/Algal cell antibodies / Conifers

product information

V-ATPase c subunit is located in vacuole and is involved in ATP synthesis cuopled proton transport. This protein is coded by ATVHA-C3 gene. Alternative names: AT4g34720/T4L20_300

KLH-conjugated synthetic peptide derived from Arabidopsis thaliana V-ATPase subunit c,Q6IDA4

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.

ELISA (ELISA), western blot (WB)

collection of antibodies to vacuolar proteins

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

AS09 503 | anti-V-ATPase, B | vacuolar ATP synthase subunit beta

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

Antibody will only work properly on purified vacuolar membranes

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 membranes are used, one µg load per well should be sufficient. Subunit c is one of most hydrophobic proteins (can be dissolved in organic solvent such as a mixture of chloroform/methanol solution). It is prone to aggregation even in the presence of SDS. Therefore, before loading on the gel membrane fractions should be incubated in buffer containing 2 % SDS at 60° or 70°C for 10 min or at 25°C for 30 min.

Protocol of isolation of plant vacuolar membranes can be found here.

application information

1: 8000 (ELISA), 1: 2000 with standard ECL (WB)

16 | 16 kDa (Arabidopsis thaliana)

Arabidopsis thaliana, Raphanus sativus

dictos including: Cucumis sativus, Gossypium mexicanum, Phaseolus aureus, Rcicinus communis, monocots including: Oryza sativa, Triticum aestivum, Zea mays, trees: Picea sitchensis, Populus trichocarpa

no confirmed exceptions from predicted reactivity known in the moment

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.

Yoshihiro et al. (2006) Immunochemical analysis of aquaporin isoforms in Arabidopsis suspension-cultured cells. Cells. Biosci.Biotechnol. Biochem. 70: 980-987.

application example

 1 µg, 5 and 10 µg of vacuolar membrane fraction/lane from Raphanus sativus  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 c subunit antibodies (AS09 468, 1:1000, 1h) and secondary anti-rabbit (1:2000, 1 h). All steps were performed in RT with agitation.