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

Product no: AS20 4369

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

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  • Product Info
  • Immunogen:

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

    Host: Rabbit
    Clonality: Polyclonal
    Purity: Immunogen affinity purified serum in PBS pH 7.4.
    Format: Lyophilized
    Quantity: 50 g
    Reconstitution: For reconstitution add 50 l of sterile water
    Storage: Store lyophilized/reconstituted at -20C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
    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
  • 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
  • Additional information (application):

  • 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.
  • Protocols
  • Method for isolation of plant vacuolar membranes

    method for vacuolar membrane isolation

    Courtesy of Dr. Masayoshi Maeshima

     

     

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Buy 2 items of this product for 239.00 €/items
Buy 3 items of this product for 217.00 €/items
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