V-ATPase, c | Vacuolar H+-ATPase, subunit c (16 kDa)

AS09 468 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, M. crystallinum, N. tabacum

Product Information

Immunogen

KLH-conjugated synthetic peptide derived from Arabidopsis thaliana V-ATPase subunit c UniProt: Q6IDA4, TAIR: AT2G16510

Host Rabbit

Clonality Polyclonal

Purity Affinity purified serum in PBS, pH 7.4

Format Lyophilized

Quantity 50 µg

Reconstitution For reconstitution add 50 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)

Recommended dilution 1 : 1000 (WB)

Expected | apparent MW

16 | 16 kDa (Arabidopsis thaliana)

Reactivity

Confirmed reactivity Arabidopsis thaliana, Mesembryanthemum crystallinum, Nicotiana tabacum

Predicted reactivity

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

Not reactive in Algae

Application examples

Application examples Application example

Following samples were analyzed: MW markers (1), Arabidopsis thaliana tonoplast (2), Arabidopsis thaliana plasma membrane (3), Arabidopsis thaliana microsomes (4), Arabidopsis thaliana total protein (5), Mesembryanthemum crystallinum tonoplast (6), Mesembryanthemum crystallinum plasma membrane (7), Mesembryanthemum crystallinum microsomes (8), Nicotiana tabacum microsomes (9), Brassica napus microsomes (10). 15 µg of the indicated protein, extracted according to Vera-Estrella et al. (2012) was separated on 12% SDS-PAGE and blotted 1.15h to PVDF using tank transfer. Blots were blocked with for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 O/N at RT with agitation. The antibody solution was decanted and the blot was washed 3X for 15 min min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera AS09 602 ) diluted to 1:50 000 in for 2h at RT with agitation. The blot was washed as above and developed using chemiluminescent substrated and recorder using the LiCOR c-DIGIT personal imager.

Courtesy Dr. Bronwyn Barkla. UNAM, Mexico

Additional information

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.

Protein or membrane sample should be treated at 70°C for 10 min before loading on the gel.

Related products

Collection of antibodies to vacuolar proteins
AS09 467 | Anti-V-ATPase subunit A | vacuolar H+-ATPase, rabbit antibodies
AS09 503 | Anti-V-ATPase, B | vacuolar ATP synthase subunit beta, rabbit antibodies
ASS09 497 | Anti- V-ATPase subunit D |V-type proton ATPase subunit D, rabbit antibodies
AS07 213 | Anti-V-ATPase subunit E of tonoplast H+ATPase, rabbit antibodies
AS09 499 | Anti-V-ATPase subunit H |V-type proton ATPase subunit H, rabbit antibodies

Plant protein extraction buffer

Secondary antibodies

Background

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

Product citations

Selected references Vera-Estrella et al. (2017). Cadmium and zinc activate adaptive mechanisms in Nicotiana tabacum similar to those observed in metal tolerant plants. Planta. 2017 Apr 28. doi: 10.1007/s00425-017-2700-1.
Barkla et al. (2016). Single-cell-type quantitative proteomic and ionomic analysis of epidermal bladder cells from the halophyte model plant Mesembryanthemum crystallinum to identify salt-responsive proteins. BMC Plant Biol. 2016 May 10;16(1):110. doi: 10.1186/s12870-016-0797-1.