Ricin Antibody | RA999

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Ricin Antibody | RA999 gallery image 1

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Mouse anti ricin A chain antibody, clone RA999 used for the detection of wild type ricin following incubation with HEK293 cells in vitro by western blotting under non-reducing conditions.
Image caption:
DHF ricin and IHF ricin show decreased binding to the cell surface and increased degradation in low pH vesicles. (A) HEK293 cells after 3 hours incubation with wild-type ricin, IHF ricin or DHF ricin run on SDS/PAGE under non-reducing conditions. A representative membranes after Western blot with anti-RTA antibodies are shown. Western blots with anti-tubulin antibodies were performed to show equal loading control. Data from three independent experiments are presented. RTA wt is marked as 1, other results are relative to this control. (B) Cells were incubated with wild-type ricin, IHF ricin or DHF ricin for 30 min at 0°C. Binding was measured as described in Methods. A representative membranes after Western blot with anti-RTA antibodies are shown. Western blots with anti-tubulin antibodies were performed to show equal loading control. Data from three independent experiments are presented. RTA wt is marked as 1, other results are relative to this control. (C) Coomasie Blue-stained 12% SDS/PAGE gels showing the stability of 500 ng of wild-type ricin, and modified ricin IHF and DHF after 30 min incubation at 0°C. (D) Cells were treated with or without bafilomycin A1 (0.1 µM) and with wild-type ricin, IHF ricin or DHF ricin. The amounts of ricin that remain in the cell after degradation was analyzed after SDS-PAGE run under non-reducing conditions (see Methods). Representative membranes after Western blot with anti-RTA antibodies are shown. Western blots with anti-tubulin antibodies were performed to show equal loading control. Data from three independent experiments are presented. Ricin wt, ricin IHF and ricin DHF in each graph are marked as 1, other results are relative to this control. All degraded forms of ricin were analyzed. (E) Tyrosine sulfated wild-type ricin sulf-1, and modified ricin sulf-1: IHF or DHF in cells after 3 hours incubation with Na235SO4 and further 3 h incubation with the toxins, run on SDS/PAGE under non-reducing conditions. Control experiments revealed equal total sulfation of proteins in cell lysates. A representative autoradiogram is shown.

From: Sokołowska I, Piłka ES, Sandvig K, Węgrzyn G, Słomińska-Wojewódzka M. Hydrophobicity of protein determinants influences the recognition of substrates by EDEM1 and EDEM2 in human cells.
BMC Cell Biol. 2015 Feb 6;16:1.

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  • Mouse anti Ricin A Chain
(Rated 0.0 out of 5 based on 0 customer reviews)
  • Product Type
    Monoclonal Antibody
  • Clone
    RA999
  • Isotype
    IgG1
1 Formats Available
    Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
    MCA2865E, WBdatasheet pdfdatasheet pdf0.2 mg
    MCA2865
    Summary
    Secondary Antibodies
    Negative Isotype Controls
    Useful Reagents
    Positive Controls
    Histology Controls
    More Images
    References
    Reviews
    -
    • Mouse anti Ricin A Chain antibody, clone RA999 recognizes the A chain of the ricin toxin (RCA60), a 60kDa glycoprotein. Mouse anti Ricin A Chain antibody, clone RA999 also cross-reacts with RCA120.

      The ricin toxin consists of two chains, A and B, covalently linked by a disulfide bond. The A chain acts as a glycosidase that removes an adenine residue from the 28S rRNA. The area from which the adenine is removed is essential for binding of elongation factors, destroying it leaves the ribosome inactive and unable to support protein synthesis. The A chain can inactivate a few thousand ribosomes per minute, much faster than the cell can replace them, meaning that a single ricin A chain can kill a cell.

      The B chain binds to complex carbohydrates on the cell surface to facilitate entry of the A chain into the cell. Without the B chain, the A chain has very low toxicity.Ricin is easily extracted from the castor bean (Ricinus communis), and the lethal dose for humans is very small. It is toxic when inhaled, ingested or injected and there is no known antidote, although a vaccine has been developed. Treatment is symptomatic and supportive, with long term organ damage likely in survivors. The ease of extraction and distribution makes it a potential candidate for biological warfare although it is less stable than other agents, such as anthrax.
    • Intended Use
    • Target Species
      Plants
    • Product Form
      Purified IgG - liquid
    • Reconstitution
    • Preparation
      Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
    • Preservative Stabilisers
      0.09%Sodium Azide (NaN3)
    • Immunogen
      Full length Ricin A (RCA60) from Ricinus communis.
    • Purity
    • Approx. Protein Concentrations
      IgG concentration 1.0mg/ml
    • Reagents In The Kit
    • Preparing The Antibody
    • Test Principle
    • Buffer Solution
      Phosphate buffered saline
    • Storage
      Store at +4oC or at -20oC if preferred.
      Storage in frost-free freezers is not recommended.
      This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
    • Shelf Life
      18 months from date of despatch.
    • GO Terms
      pathogenesis
      negative regulation of translation
      nucleotide binding
      sugar binding
      rRNA N-glycosylase activity
      defense response
    • UniProt
    • Entrez Gene
    • Acknowledgements
    • Regulatory
      For research purposes only
    • This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.

    • Application NameYesNoMin DilutionMax Dilution
      ELISA
      Western Blotting

    • Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
    • Technical Advice
    • Recommended Protocol
    • ELISA
    • Immunohistology
    • Histology Positive Control Tissue
    • Immunofluorescence
    • Western Blotting
    • Instructions For Use

    Additional Ricin Antibody Formats

    Formats Clone Applications Sizes available
    Ricin Antibody : Purified RA999 E, WB 0.2 mg
    • Copyright © 2016 Bio-Rad Antibodies (formerly AbD Serotec)

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      Recommended Negative Isotype Control

        Useful Reagents

          Recommended Positive Controls

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              References

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