VDAC1 | voltage-dependent anion-selective channel protein 1

320 €

AS07 212 | clonality: polyclonal | host: rabbit | reactivity: A.thaliana, di and monocots,| cellular [compartment marker] of mitochondrial outer membrane

PRODUCT INFORMATION IN PDF

product information

background

VDAC1 protein (called also Synonymes: At3g01280, outer mitochondrial membrane protein porin 1, T22N4_9, T22N4.9, VDAC 1, Voltage-dependent anion-selective channel protein 1, voltage-gated ion-selective channel) forms a channel through the cell membrane for diffusion of small hydrophilic molecules. Evolutionary origin of VDAC protein is not clear and their structure and properties are quite different making those proteins only conceptually like porins (Clausen et al. 2004).

immunogen

KLH-conjugated peptide conserved in all known higher plant VDAC1 proteins including Arabidopsis thaliana with the locus name: At3g01280

antibody format

rabbit

polyclonal

affinity purified serum, in PBS pH 7.4

lyophilized

quantity

200 µg

for reconstitution add 200 µl of sterile water.

storage

store lyophilized/reconstituted at -20°C; once reconstituted 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 lyophilized material adhering to the cap or sides of the tubes.

tested applications

western blot (WB), Blue-native (2D BN/SDS-PAGE), immunolocalization (IL)

related products

AS04 054 | anti-AOX1/2 rabbit antibody, marker of mitochondrial inner membrane

AS06 203A | anti-Idh rabbit antibody, marker of mitochondrial matrix

collection of antibodies to other mitochondrial proteins

additional information

cellular [compartment marker] of mitochondrial outer membrane

application information
recommended dilution		1:5000 on 2-30 µg of protein/lane with standard ECL (WB), 1: 500 (IL)
expected \| apparent MW		29 kDa (for Arabidopsis thaliana)
confirmed reactivity		Arabidopsis thaliana, Beta vulgaris, Papaver sp. pollen tubes (IL), Spinacia oleracea, Physcomitrella patens,
predicted reactivity		dicots including Nicotiana tabacum, Plantago major, Ricinus communis,monocots including Triticum aestivum,
not reactive in		Chlamydomonas reinhardtii, Glycine max, Zea mays, diatoms
additional information		Amount of mitochondrial fraction detected by anti-VDAC1 antibody was from 2-10 µg. Immunolocalization method description and images are available here Blue-native (2D BN/SDS-PAGE) methodology is described in Piechota et al. 2010
selected references		Alcántar-Aguirre et al.(2013).ATP produced by oxidative phosphorylation is channeled toward hexokinase bound to mitochondrial porin (VDAC) in beetroots (Beta vulgaris). Planta, March 17. Zhou et al. (2012). UBIQUITIN-SPECIFIC PROTEASE16 Modulates Salt Tolerance in Arabidopsis by Regulating Na+/H+ Antiport Activity and Serine Hydroxymethyltransferase Stability. Plant Cell, dec. 24. Lang, E.G.E., S.J. Mueller, S.N.W. Hoernstein, J. Porankiewicz-Asplund, M. Vervliet-Scheebaum, R. Reski (2010). Simultaneous isolation of pure and intact chloroplasts and mitochondria from moss as basis for sub-cellular proteomics. Plant Cell Reports, DOI: 10.1007/s00299-010-0935-4. (open source)

application example

western blot using anti-VDAC1 antibodies

10 μg of mitochondrial fraction from Arabidopsis thaliana and 25 μg of Arabidopsis thaliana leaf extract were separated on 10% gel and blotted on nitrocellulose membrane using wet transfer (0.22% CAPS, pH 11). Filters where blocked (1.5h) in 5% milk in TBST (1X TBS, 0,1% Tween 20), incubated with 1: 5000 anti-VDAC1 antibodies (2h in TBST) followed by incubation with 1: 10 000 secondary anti-rabbit (1h) HRP-coupled antibodies and visualized with standard ECL on Kodak autoradiography film for 15-60 s. Mitochondria were isolated as described by Urantowka et al. (Plant Mol Biol, 2005, 59:239-52). Mitochondrial pellets were suspended in 1X Laemmli buffer (5% beta-mercaptoetanol, 3.7% glycerol, 1.1% SDS, 23 mM Tris- HCl pH 6.8, 0.01% bromophenol blue), heated (95°C, 5 min.) and centrifuged (13 000rpm, 1 min.). Leaf extracts were prepared as described by Martinez-Garcia et al. (Plant J., 1999, 20:251-7).

Courtesy Dr. Janusz Piechota, Wrocław University, Poland

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