IKP

Api g 1 | Major allergen Api g 1, isoallergen 2

Product no: AS19 4335

AS19 4335 | Clonality: Polyclonal  |  Host: Rabbit  | Reactivity: Apium graveolens

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  • Product Info
  • Immunogen: Recombinant Apium graveolens Major allergen Api g 1, isoallergen 2 protein, amino acids: 1-159, UniProt:P92918
    Host: Rabbit
    Clonality: Polyclonal
    Purity: >95%, Protein G purified to a total immunoglobulin G fraction.
    Format: Liquid
    Quantity: 50 g
    Storage: Store at -20C or -80C, avoid repeated freeze-thaw cycles. 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: ELISA (ELISA)
    Recommended dilution: to be determined by end user
  • Reactivity
  • Confirmed reactivity: Apium graveolens
  • Application Examples
  • Western blot using anti-Api g 1 major allergen antibodies

    Samples:
    1 – 12 µg celery (Apium graveolens) leaves, proteins extracted with 10% TCA/Acetone
    2 – 12 µg (Apium graveolens) stems, proteins extracted with 10% TCA/Acetone
    3- 12 µg (Apium graveolens) leaves, proteins extracted with 50mM Tris-HCl, pH7.4; 150 mM NaCl, protease inhibitors, followed by protamine sulfate depletion (0.1% for RuBisCO removal) and precipitated with 10% TCA/Acetone
    4- 12 µg (Apium graveolens) leaves, proteins extracted with 50mM Tris-HCl, pH7.4; 150 mM NaCl, protease inhibitors
    MW marker (kDa): PageRuler™ Prestained Protein Ladder (Thermo Fisher Scientific)

    12 µg/well of total protein extracted freshly from celery (Apium graveolens) with 10% TCA/ Actetone (samples 1 and 2) and with 50mM Tris-HCl, pH7.4; 150 mM NaCl, protease inhibitors, followed by protamine sulfate depletion (0.1% for RuBisCO removal) and precipitated with 10% TCA/Acetone (samples 3 and 4) and denatured in Laemmli buffer at 70°C for 5 min. The samples were separated on 10 % SDS-PAGE and blotted 1h to nitrocellulose using wet transfer. Blot was blocked with 2 % milk TBS-T at 4°C/ON without agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 for 1h/RT with agitation in TBS-T 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. Blot was incubated in Agrisera matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, AS09 602) diluted to 1:25 000 in TBS-T for 1h/RT with agitation. The blot was washed as above and developed for 5 min with Agrisera ECL SuperBright. Exposure time was 60 seconds.

    Courtesy of Yordan Muhovsky, Walloon Agricultural Research Centre CRA-W · Department of Life Sciences, Belgium

  • Additional Information
  • Additional information: Preservative: 0,03% Proclin 300, Preparation contains: 50% Glycerol, 10 mM PBS, pH 7,4
  • Background
  • Background: Major allergen Api g 1, isoallergen 2 (Allergen Api g 1.0201) (allergen Api g 1)
  • Product Citations
  • Selected references: 38221900
  • Protocols
  • ELISA protocol

    Protocols to work with plant and algal protein extracts

    Agrisera Educational Poster Collection
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