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 tonoplast proteins

AS09 492 | anti-TIP1;1, TIP1;2 | tonoplast intrinistic protein 1-1, 1-2 (gamma)

AS09 493 | anti-TIP1;1, TIP1;2 | tonoplast intrinistic protein 1-1, 1-2 (gamma)

AS09 510 | anti-TIP2;1 | tonoplast intrinistic protein 2-1

AS09 511 | anti-TIP2;2 | tonoplast intrinistic protein 2-2 (C-terminal)

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.

42 kDa dimer is also detected by those antibodies.

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

application information
recommended dilution		1: 8000 (ELISA), 1: 1000 with standard ECL (WB)
expected \| apparent MW		24.9 \| 23 kDa
confirmed reactivity		Raphanus sativus
predicted reactivity		Raphanus sativus
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		to be added when available

application example

crude membrane of yeast cells expressing recombinaTIP2;1 (RsTIP2;1) and TIP1;1 (RsTIP1;1) proteins 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-TIP2;1 antibodies (AS09 494, 1:2000, 1h) and secondary anti-rabbit (1:2000, 1 h). All steps were performed in RT with agitation.
Left panel: antibody incubacted without peptide, right panel with peptide used to elicit this antibody (peptide competition assay)

western blot using anti-TIP2;1 antibodies

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