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V-ATPase, a3 | vacuolar H+-ATPase subunit a isoform 3

AS20 4369 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

V-ATPase, a3 | vacuolar H+-ATPase subunit a isoform 3 in the group Antibodies for Plant/Algal  / Membrane Transport System / Vacuolar membrane at Agrisera AB (Antibodies for research) (AS20 4369)

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Product Information

Immunogen

KLH-conjugated synthetic peptide derived from Arabidopsis thaliana V-ATPase subunit a3, UniProt: Q8W4S4-1 , TAIR:  At4g39080

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

92.8 | >100 kDa (Arabidopsis thaliana)

Reactivity

Confirmed reactivity Arabidopsis thaliana
Predicted reactivity Brassica campestris, Brassica oleracea, Brassica rapa, Capsella rubella, Eutrema salsugineu, Noccaea caerulescens
Species of your interest not listed? Contact us
Not reactive in No confirmed exceptions from predicted reactivity are currently known.

Application examples

Application examples Aplication example

Westen blot using anti-V-ATPase, a3 antibodies

30 µg of microsomal membranes were extracted freshly from 5-day-old Arabidopsis thaliana seedlings grown in liquid culture with a buffer containing 350mM sucrose, 70mM Tris-HCl pH8, 10% glycerol, 3mM Na2EDTA, 0.15% BSA, 1.5% PVP-40, 4mM DTT, 1x Roche completeTM Protease Inhibitor and denatured with 1x Lämmli buffer at 50°C for 10 min. Proteins were separated on 7,5 % SDS-PAGE and blotted 1h to PVDF (pore size of 0.2 µm) using wet transfer. Blot was blocked with 3% milk for 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 in 2%BSA in TBS-T (0.05%) for ON/4°C with agitation. The antibody solution was decanted and the blot was rinsed briefly with TBS-T, then washed twice for 5 min in TBS-T at RT with agitation. Blot was incubated in Agrisera matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:25 000 in 3% milk in TBS-T for 2h/RT with agitation. The blot was washed as above and developed for 2 min with chemiluminescent detection reagent. Exposure time was 100 seconds.

expected band sizes: 1 - marker as indicated in the picture 2 - endogenous VHA-a3 at ~93kDa 3 - endogenous VHA-a3 at ~93kDa 4 - no band since it is the mutant (It also tells us that the AB is not recognizing VHA-a2 which is very similar to VHA-a3) 5 - no band since it is the mutant (It also tells us that the AB is not recognizing VHA-a2 which is very similar to VHA-a3) 6 - endogenous VHA-a3 at ~93kDa (It also tells us that the AB is not recognizing VHA-a1(-GFP) which is similar to VHA-a3) 7 - endogenous VHA-a3 at ~93kDa and VHA-a3-GFP at ~120kDa

Courtesy of Upendo Lupanga, Centre of Organismal Studies, University of Heidelberg, Germany

Additional information

Related products

Related products AS14 2822 | Anti-V-ATPase, a1 | vacuolar H+-ATPase subunit a isoform 1, rabbit antibodies

collection of antibodies to other vacuolar membrane proteins

Background

Background V-ATPase subunit VHA-a3 The protein is essential component of the vacuolar proton pump (V-ATPase), a multimeric enzyme that catalyzes the translocation of protons across the membranes and required for assembly and activity of the V-ATPase. Involved in vacuolar nutrient storage (e.g. accumulation and storage of nitrate) and in tolerance to some toxic ions (e.g. zinc ions sequestration in vacuoles).

Alternative names: V-type proton ATPase 95 kDa subunit a isoform 3, V-ATPase 95 kDa isoform a3, Vacuolar H(+)-ATPase subunit a isoform 3, Vacuolar proton pump subunit a3, Vacuolar proton translocating ATPase 95 kDa subunit a isoform 3.

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