Anti-Ricin (RTA subunit), clone MMA

Product no: AS09 648

AS09 648 | Clonality: Polyclonal | Host: Mouse | Reactivity: Ricinus communis

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Delivery:  7-10 business days
  • Product Info
  • Sub class: IgG1

    Ricin, RTA subunit

    Host: Mouse
    Clonality: Monoclonal
    Purity: Purified IgG in PBS pH 7.4 with mannitol.
    Format: Lyophilized
    Quantity: 200 µg
    Reconstitution: For reconstitution add 200 µl of sterile water
    Storage: Store at -70°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. , For storage at 4°C sodium azide can be added
    Tested applications: ELISA (ELISA), Immunohistochemistry (IHC)
    Recommended dilution: The optimal working dilution should be determined by the investigator, for specific application
    Expected | apparent MW: 29.8 kDa
  • Reactivity
  • Confirmed reactivity: Ricinus communis
    Predicted reactivity: Ricinus communis
    Not reactive in: No confirmed exceptions from predicted reactivity are currently known
  • Additional Information
  • Additional information:

    Antibodies have been purified by affinity chromatography on Protein G from tissue culture supernatant and lyophilized from solution containing: mannitol 1%, dextran 1%, NaCl 100mM, sodium phosphate 10 mM, pH 7.5. If required, mannitol can be removed either by dialysis or by chromatography using mini columns Sephadex G50 fine. 

    The concentration can be measured by absorbance at 280 nm (1.35 for 1 mg/ml) or by Bradford method

    Ricin detection limit is 5 ng/ml, signal-to-noise ratio is 15-50 up to 100 ng/ml

    Epitope recognized by this antibody is located on the surface of RTA subunit and not accessible in intact ricin molecule. 

    Suggested protocol can be found here.

    Additional information (application): Detected epitope is located on the surface of RTA subunit not accssible in intact ricin molecule, This antibody does not cross-react to whole ricin molecule
  • Background
  • Background:

    Ricin is a protein found in castor bean (Ricinus communis) acting as protein synthesis inhibitor. It is a toxin involved in plant defence. A chain of ricin can inactivate few thousands of ribosomes per minute. B chain facilitates the entry of A chain into the cell. 

  • Protocols

    Materials and Reagents
    1. 96-well plates
    2. Suitable plate reader (depending on the detection system)
    3. Plate shaker with temperature control
    4. Capture antibody (#177 YO Proteins)
    5. Detection antibody (conjugate) (enquire)
    6. Washing solution (PBS, 0.025% Tween 20, 1% D-lactose)
    7. Solution for blocking and dilution (PBS, 0.025% Tween 20, 1% D-lactose,
    0.1% BSA)
    8. TMB reagent
    9. Sulfuric acid 10%

    1. Immobilisation of capture antibody
    Dissolve capture antibody in PBS at 5 μg/ml. Add 100 μl into each well. Incubate 2
    hours at 37°C. Wash twice with 300 μl washing solution.
    2. Blocking
    Add 250 ml blocking solution. Incubate 1 hour at 37°C. Wash with washing solution
    twice 300 μl.
    3. Application of control and samples.
    Add 100 μl to each well: antigen (ricin) diluted from 0.5 to 100 ng/ml, sample serially
    diluted. Incubate 1 hour at 37°C. Washing 3 times 300 μl.
    4. Detection antibody (example based on biotin conjugate)
    Add 100 μl of biotin-conjugated antibody (1 μg/ml). Incubate 1 hour at 37°C.
    Washing 3 times 300 μl (not shorter than 5 min each).
    5. Streptavidin-HRP conjugate
    Add 100 μl of Streptavidin-HRP conjugate (diluted 10000X). Incubate 1 hour at
    37°C. Washing 5 times 300 μl (not shorter than 5 min each).
    6. Development
    Add 100 μl of development TMB solution. Incubate 10 min at 37°C. Stop reaction
    with 10 μl 10% sulfuric acid. Read OD at 450 nm

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