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V-ATPase, a1 | vacuolar H+-ATPase subunit a isoform 1

AS14 2822 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

V-ATPase, a1 | vacuolar H+-ATPase subunit a isoform 1 in the group Plant/Algal Antibodies / Membrane Transport System / Vacuolar membrane at Agrisera AB (Antibodies for research) (AS14 2822)

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product information
Background V-ATPase subunit VHA-a1 is a subunit of the membrane-integral Vo subunit. VHA-a1 target the V-ATPase enzyme to the trans-golgi network/earl endosome (TGN/EE) in Arabidopsis thaliana. This enzyme is involved in acidification process of various compartements of eucaryotic cells. The protein is coded by VHA-a1 gene AT2G28520. Alternative names: vacuolar H(+)-ATPase subunit a1, V-ATPase 93 kDa subunit.
Immunogen

KLH-conjugated synthetic peptide derived from Arabidopsis thaliana V-ATPase subunit A, O23654, At1g78900

Host Rabbit
Clonality Polyclonal
Clone
Purity Affinity purified serum in PBS, pH 7.4
Format Lyophilized
Quantity 100 µg
Reconstitution For reconstitution add 100 ĩl 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)
Related products

collection of antibodies to other vacuolar membrane proteins

Plant protein extraction buffer

Secondary antibodies

Additional information

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

application information
Recommended dilution 1 : 1000 (WB)
Expected | apparent MW

93 kDa (Arabidopsis thaliana)

Confirmed reactivity Arabidopsis thaliana
Predicted reactivity Capsella rubella, Cucumis sativus, Erythranthe guttata , Glycine soja, Lupinus angustifolius, Morus notabilis, Phaseolus vulgaris , Vitis vinifera
Not reactive in No confirmed exceptions from predicted reactivity are currently known.
Additional information

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

Selected references

To be added when available, antibody released in April 2017. 


aplication example

western blot using anti-VHa1 antibodies

10 μg of microsomal membranes were isolated from 6-days-old etiolated (dark-grown) Arabidopsis thaliana seedlings with Extraction buffer (450mM sucrose, 50mM HEPES pH7.5, 5mM MgCl2, 1mM DTT, 1x protease inhibitor). Proteins were denaturated in SDS sample buffer for 5 min at 95°C. Proteins were separated on a 10% SDS-PAGE and blotted 1h to a nitrocellulose membrane using tank transfer. Blots were blocked with 4% dry milk in TBS-T for 2h at room temperature (RT) with agitation. Blot was incubated in the primary antibody (#AS14 2822) at a dilution of 1:1000 in blocking buffer for 16h with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed 3 times for 15 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 1:25 000 in blocking buffer for 2h at RT with agitation. The blot was washed as above and developed for 3 min with ECL according to the manufacturer's instructions peqlab AceGlow Kit & INTAS digital imager. Exposure time was 3 min.

Courtesy of Fabian Fink, University Heidelberg, Germany

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