Agrisera/TIP1 | Tonoplast intrinsic protein 1-1/AS09 482/
| | product information | | Background | | TIP1;1 is a water channel protein needed for the transport of water, diffusion of amino acids and peptides. Alternative name: Tonoplast intrinsic protein 1-1, gamma-tonoplast intrinsic protein, Gamma-TIP, aquaporin TIP, Tonoplast intrinsic protein, root-specific RB7 | | Immunogen | | KLH-conjugated synthetic peptide derived from Arabidopsis thaliana P25818, At2g36830 | | Host | | Rabbit | | Clonality | | Polyclonal | | Clone | | | | Purity | | Serum | | Format | | Lyophilized | | Quantity | | 100 µl | | Reconstitution | | For reconstitution add 200 µl of sterile water. | | 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 tonoplast proteins AS09 493 | anti-TIP1;1, TIP1;2 | tonoplast intrinistic protein 1-1, 1-2 (gamma) AS09 494 | anti-TIP2;1 (delta) | Delta-VM23 AS09 510 | anti-TIP2;1 | tonoplast intrinistic protein 2-1 AS09 511 | anti-TIP2;2 | tonoplast intrinistic protein 2-2 (C-terminal) Plant protein extraction buffer Secondary antibodies | | 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. TIP1;1 usually shows a faint band of dimeric form at 42 kDa in addition to the major monomoer band of 23 kDa even in the presence of SDS. Protocol for isolation of plant vacuolar membranes can be found here. | |
| application information |
| Recommended dilution | | 1: 8000 (ELISA), 1: 2000 with standard ECL (WB) |
| Expected | apparent MW | | 25.6 | 24 kDa (Arabidopsis thaliana) |
| Confirmed reactivity | | Arabidopsis thaliana |
| Predicted reactivity | | dictos including: Brassica oleracea, Gossypium mexicanum, Pisum sativum, Ricinus communis, Vitis vinifera, monocots including: Oryza sativa, trees: Picea abies, Popoulus trichocarpa |
| 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 | | Ishikawa et al. (2005). Novel type aquaporin SIPs are mainly localized to the ER membrane and show cell-specific expression in Arabidopsis thaliana. FEBS Lett. 25:5814-5820. |
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application example
1 µg and 10 µg of crude membrane fraction/lane from Arabidopsis thaliana 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-TIP1;1 antibodies (AS09 482, 1:1000, 1h) and secondary anti-rabbit (1:2000, 1 h). All steps were performed in RT with agitation.
TIP1;1 usually shows a faint band of dimeric form at 42 kDa in addition to the major monomoer band of 23 kDa even in the presence of SDS.
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