ASTM D6499-12 Antigenic Protein in Natural Rubber and its Products

ASTM D6499-12 Antigenic Protein in Natural Rubber and its Products

ASTM D6499-12 covers an immunological technique to decide how much antigenic protein is in regular elastic. This test utilizes rabbit antisera for regular elastic latex (NRL) proteins. The amount of protein is estimated at microgram to milligram levels.

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    Using an inhibitory format, this immunoassay method, ASTM D6499-12, quantifies the quantity of antigenic latex proteins in solution. ASTM D6499 depicts an immunological strategy for quantitating normal elastic latex proteins. 


    In ASTM D6499-12, the latex sample is suspended for two hours in fluid support. The concentrate is recuperated, and the antigen levels are measured utilizing restraint Enzyme-Linked Immuno Sorbent Assay (ELISA) technology. The test extricates are blended in with neutralizer explicit for NRL protein in a weakening plate. Following concise hatching to consider immunizer acknowledgment of the pertinent NRL antigens, the combination is added to the immobilized antigen in the examine plate. Next, the enzyme-substrate is added, and the enzyme’s reaction to the substrate results in a color change.

    Apparatus and Specimen

    In ASTM D6499-12, 96-Well Microtiter Assay Plate, Dilution Plate, Multichannel Pipettors, Analytical Balance, Centrifuge, (capable of 1000 × g) and tubes, An Incubator, capable of regulating the temperature at ~37°C, Microtiter Plate Reader, optional computer for data analysis, and ELISA Plate Sealing Tape or Plastic Lids are required.

    Gloves or other rubber product extracts may be included in the samples, which were collected to determine latex protein levels.


    The absorbance readings of the test samples are changed over to μg protein/mL. The outcomes with a bend fitting PC program that utilizes either a quadratic polynomial guess, a spline estimation, or a Semi-log change of the absorbance information are obtained.


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