Anti-PDHE1-A | Pyruvate dehydrogenase E1 component subunit alpha-1, mitochondrial

Product no: AS23 4974

AS23 4974  |  Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity: Arabidopsis thaliana

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  • Product Info
  • Immunogen: KLH-conjugated, unique peptide derived from Arabidopsis thaliana PDHE1-A, UniProt: P52901, TAIR: AT1G59900 

    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 : 500 - 2000 (WB)
    Expected | apparent MW: 43.1 | 39 kDa (signal peptide is removed)
  • Reactivity
  • Confirmed reactivity: Arabidopsis thaliana
    Predicted reactivity: Brachypodium distachyon, Brassica napus, Cannabis sativa, Capsicum annuum, Cucumis sativus, Glycine max, Gossypium sp., Hordeum vulgare, Malus domestica, Manihot esculenta, Medicago truncatula, Nicotiana tabacum, Oryza sativa, Pisum sativum, Populus sp., Solanum lycopersicum, Solanum tuberosum, Sorghum bicolor, Spinacia oleracea, Theobroma cacao, Triticum sp., Vitis vinifera, Zea mays
    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-PDHE1-A antibodies

    Samples:
    1 - 40 µg of Arabidopsis thaliana Col-0 whole leaf extract.
    2 - 40 µg of Arabidopsis thaliana mutant iar4 (AT1G24180), Arabidopsis thaliana pyruvate dehydrogenase E1a-like subunit.
    3 - 40 µg of Arabidopsis thaliana mutant iar4l-1 (AT1G59900), Arabidopsis thaliana pyruvate dehydrogenase E1a subunit.
    4 - 40 µg of Arabidopsis thaliana mutant iar4l-2 (AT1G59900), Arabidopsis thaliana pyruvate dehydrogenase E1a subunit.
    5 - 40 µg of Arabidopsis thaliana mutant SALK_004367C (AT5G50850), Arabidopsis thaliana pyruvate dehydrogenase E1 component subunit beta-1.

    40 µg/well of total protein extracted freshly from Arabidopsis thaliana whole rosette leaves.   Exact buffer components were: 50 mM Tris-HCl (pH 7.5), 150 mM NaCl, 0.5 mM EDTA, 10% (v/v) glycerol, 1% (v/v) Nonidet P-40 (NP-40), 1% (w/v) deoxycholate, 0.1% (w/v) SDS, 1 × Complete protease inhibitor cocktail (Roche), 1 mM PMSF, and denatured with 5xSDS sample buffer (300 mM Tris-HCL(pH 6.8), 10% SDS, 0.1% Bromophenol, 50% Glycerol, 500 mM DTT) at 95°C/5 min.  Samples were separated in the RT on 10 % SDS-PAGE and blotted for 7 min to nitrocellulose (pore size of 0.2 um), using:  iBlotTM Dry Blotting System (Invitrogen) in the RT. Blot was blocked with 5% milk 5  for: 3h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1:  500 for ON/4°C with agitation in 2% milk. The antibody solution was decanted, and the blot was rinsed briefly twice, then washed 5 times for 5 min in PBS-T at RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1: 5000 in 2% milk for 1h/RT with agitation. The blot was washed as above and developed with a following chemiluminescent detection reagent: AS16 ECL-N-10 Agrisera Bright. Exposure time was 1 minute.

    To increase signal/noise ratio, primary antibody can be used in a dilution of 1: 1000 1h/RT incubation. 

    Courtesy of Dr. Mengping Li, Department of Botany and Plant Biology, UNIGE, Switzerland

  • Background
  • Background: PDHE1-A (Pyruvate dehydrogenase E1 component subunit alpha-1) is the e1 alpha subunit of the pyruvate dehydrogenase complex (PDC), which catalyzes the overall conversion of pyruvate to acetyl-CoA and CO2.
  • 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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Buy 2 items of this product for 239.00 €/items
Buy 3 items of this product for 217.00 €/items
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