Anti-12S seed storage protein CRC

Product no: AS20 4403
AS20 4403 | Clonality: Polyclonal  |  Host: Rabbit |  Reactivity: Arabidopsis thaliana
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  • Product Info
  • Immunogen: Native, 12S globulin alpaha subunit purified from Arabidopsis thaliana and excised from SDS-PAGE gel. UniProt: Q96318, TAIR: At4g28520
    Host: Rabbit
    Clonality: Polyclonal
    Purity: Total IgG. Protein A purified in PBS, 50% glycerol. Filter sterilized.
    Format: Liquid at 2 mg/ml.
    Quantity: 200 µg
    Storage: Store at -20°C; 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: ELISA (ELISA), Immunolocalization (IL) using electron microscopy, Immunohistochemistry (IHC), Western blot (WB)
    Recommended dilution: Assay dependent (ELISA), 1: 50 (IL by electron microscopy), 1: 100 (IHC), 1: 3000 - 1: 10 000 (WB)
    Expected | apparent MW: 58 | 30 kDa (alpha subunit)
  • Reactivity
  • Confirmed reactivity: Arabidopsis thaliana
    Predicted reactivity: 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-12S seed storage protein CRC antibodies
    Arabidopsis thaliana dry seed extract from wilde-type (1) and atvst1-1 mutant (deffective in storage protein transport (2)  (SDS-PAGE, stained gel)  Western blot of the same samples (3,4) which were separated on 15-20 % SDS-PAGE and blotted to PVDF membrane in wet system. Blot was blocked with 3 % skim milk/TBS-T, 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 5000 in TBS-T for 1h/RT. The antibody solution was decanted and the blot was washed 4 times for 10 min in TBS-T at RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 in for 1h/RT with agitation. The blot was washed as above and developed with a chemiluminescent detection reagent, following manufacture's recommendations.
    P12S is the precursor of 12S globulin. 12S-alpha is the alpha-subunit (proteolyticall form of 12S) of 12S globulin.
  • Background
  • Background: Major 12S seed storage protein CRC (globulin) is synthesized on the endoplasmic reticulum as precursor and then transported to storage vacuoles, where it is processed at a conserved Asn-Gly peptide bond by an asparaginyl endopeptidase to produce two mature polypeptides referred to as alpha and beta subunits that are joined together by a disulfide bond. Phosphorylated in seeds on some Tyr residues in response to abscisic acid (ABA). 12S CRC protein is cleaved to alpha and beta chains. 
    Cellular localisation: vacuole.

    Alternative protein names: Cruciferin 3, Cruciferin C Legumin-type globulin Cruciferin1, Legumin-type globulin storage protein CRU1.
  • Product Citations
  • Selected references: Shirakawa et al. (2014). CONTINUOUS VASCULAR RING (COV1) is a trans-Golgi network-localized membrane protein required for Golgi morphology and vacuolar protein sorting. Plant Cell Physiol. 2014 Apr;55(4):764-72.doi: 10.1093/pcp/pct195. (Immunohistochemistry, Western blot)
    Li et al. MAG2 and three MAG2-INTERACTING PROTEINs form an ER-localized complex to facilitate storage protein transport in Arabidopsis thaliana. Plant J. 2013 Dec;76(5):781-91.doi: 10.1111/tpj.12347. (Immunolocalisation by electron microscopy, Western blot)
  • Protocols
  • Agrisera Western Blot protocol and video tutorials

    Protocols to work with plant and algal protein extracts

    Agrisera Educational Posters Collection
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