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

western blot (WB), ELISA (ELISA)

related products

collection of antibodies to tonoplast proteins

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 plant vacuolar membranes can be found here.

application information
recommended dilution		1: 1000 with standard ECL (WB), 1: 8000 (ELISA)
expected \| apparent MW		16.6 \|
confirmed reactivity		Arabidopsis thaliana
predicted reactivity		Arabidopsis thaliana
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. Manuafactured by Operon Biotechnologies.
selected references		Ide et al. (2007). Transcriptional Induction of Two Genes for CCaPs, Novel Cytosolic Proteins, in Arabidopsis thaliana in the Dark. Plant Cell Physiol. 1:54-65.

application example

5 µg (1,3) and 2.5 µg (2,5) of lysate from E.coli cells expressing CCaP1 protein/lane 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-CCaP1 antibodies (AS09 483, 1:1000, 1h) and secondary anti-rabbit (1:2000, 1 h). All steps were performed in RT with agitation.

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