AHA2 p | ATPase 2 phosphorylated, plasma membrane-type

Product no: AS22 4789

AS22 4789   | Clonality:  Polyclonal |  Host:  Rabbit | Reactivity: Amaranthus cruentus, Arabidopsis thaliana, Beta vulgaris

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  • Product Info
  • Immunogen: KLH-conjugated peptide derived from Arabidopsis thaliana AHA2 protein sequence UniProt: P19456, TAIR: At4g30190
    Host: Rabbit
    Clonality: Polyclonal
    Purity: Antigen affinity purified serum, in PBS pH 7.4
    Format: Lyophilized
    Quantity: 50 µg
    Reconstitution: For reconstitution, add 50 µl, of sterile or deionized 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 : 10 000 (WB)
    Expected | apparent MW: 104.4 | kDa 
  • Reactivity
  • Confirmed reactivity: Amaranthus cruentus, Arabidopsis thaliana, Beta vulgaris
    Predicted reactivity: Brassica oleracea, Camelina sativa, Capsella rubella, Eutrema salsugineum, Raphanus sativus

    Species of your interest not listed? Contact us
    Not reactive in: No confirmed exceptions from predicted reactivity are currently known
  • Application Examples
  • Western blot using anti AHA2 phosphorylated antibodies

    1 - Recombinant protein MBP-C terminal region of AHA2, phosphorylation treatment
    2 - Recombinant protein MBP-C terminal region of AHA2, subsequently dephosphorylated
    3 - Recombinant protein MBP, phosphorylation treatment
    4 - Recombinant protein MBP-C terminal region of AHA2, Thr947Ala, phosphorylation treatment 5- Mark: MW markers

    1 µg/well of recombinant protein produced in E. coli and purified by affinity chromatography was incubated in the reaction mixture (30 ul).  3 ul of the reaction were mixed with 6 ul Laemli 2X and denatured at 95°C for 5 min.  Samples were separated on 10% SDS-PAGE and blotted overnight (ON) to PVDF (Inmobilon®-FL) (pore size of 0.45 µm) using wet transfer. The blot was blocked with 3 % non-fat milk 2h/RT with agitation. The blot was incubated in the primary antibody at a dilution of 1:10 000 for 90 min/RT with agitation 3% non-fat milk TBS with agitation. The antibody solution was decanted, and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. The blot was incubated in a matching secondary antibody (anti-rabbit IgG horseradish peroxidase conjugated) diluted to 1: 5000 for 90 min/RT with agitation. The blot was washed as above and developed with the following chemiluminescent detection reagent: AgriseraBright. Exposure time was 2 min.

    Courtesy of Cristian Mayordomo, IBMCP, Universidad Politécnica de Valencia, Spain

    Western blot on using anti-AHA2p antibodies

    Titration of the AS22 4789: AHA2 p | ATPase 2 phosphorylated, plasma membrane-type antibody. Ten µg of protein from Amaranth stem mixed membranes (TO), or from beetroot plasma membranes either pre-phosphorylated, i.e. treated with ATP (MP+P), or not (MP* and MP-P) were blotted and probed with the indicated dilutions of antibody. Molecular mass markers (MPM) have, from top to bottom 250, 130, 100, 70 (red) and 55 kDa, respectively.

    Courtesy Dr. Luis González, CINVESTAV, Mexico

  • Background
  • Background: The plasma membrane H+ ATPase of plants and fungi generates a proton gradient that drives the active transport of nutrients by H+-symport
  • Protocols
  • Agrisera Western Blot protocol and video tutorials

    Protocols to work with plant and algal protein extracts

    Agrisera Educational Poster Collection
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