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ASTM Committee D11- Rubber and Rubber Products 'Document Sheet' for- Industry Reference Material IRM-913 NR Protein

Introduction

Industry Reference Material (IRM) 913 is the designation for the ammoniate natural rubber (NR) protein reference antigen that is used as a coating antigen and reference standard according to the procedures of D6499. Details on the evaluation of this material are found in Research Report D11-1094.

Properties

Property ASTM Designation Limits/Targets Source D6499 Ammoniated NR latex (high & low) Protein Amount D5712 Stock solution should be adjusted to 1mg protein per ml. AA Analysis RR D11-1094 The correlation coefficient of AA composition between batches should be at least 0.90 SDS-PAGE RR D11-1094 Full distribution of NR proteins (between 5 kDa and 200 kDa) Western Blot Profile RR D11-1094 Full distribution of NR antigens (between 5 kDa and 200 kDa) Inhibition ELISA RR D11-1094 Adjust antigen content based on D6499 determination

Accepted Reference Value (AR Value) The standard antigen preparation Lot 1.1 was assigned a reference value of 1000 ug/ml based on the D5712 protein determination after reconstitution in 3.3 mls of H2O.

Conc ug/ml AR-Value (%) +/- 2 Sigma Min Max Range (%) 2 83.6 3.5 80.1 87.1 7.0 1 76.3 3.5 72.8 79.8 7.0 0.5 68.0 3.5 64.5 71.5 7.0 0.25 56.8 3.5 53.3 60.3 7.0 0.13 45.8 3.5 42.3 49.3 7.0 0.06 29.9 3.5 26.4 33.4 7.0 0.03 19.0 3.5 22.5 22.5 7.0

AR Value is given as percent inhibition in the D6499 assay.

Specific Conditions Under Which the IRM 913 is to be Stored

The material is stored lyophilized and sealed under nitrogen in glass vials. The glass vials should be stored a 4ºC. The shelf life of the lyophilized protein has not been determined but should have a minimum shelf life of 10 yrs provided the 4ºC conditions are maintained. The lyophilized preparation of NRL protein is reconstituted with distilled H2O to a concentration of 1 mg/ml. Aliquot this stock solution into small polypropylene tube and store at -20ºC. Aliquots, once thawed for used in the assay, should be stored at 4ºC and used within 1 week.

Ordering IRM 913 NR Protein & IRM 914 NR Antisera These IRMs are available for purchase in the following ways:

Test Package..................................$810.00 USD/Package Includes 1 vials Antisera (IRM 914) & 15 vials NR Protein (IRM 913) Sufficient for ASTM D 6499 testing of 7500 samples Replacement

Replacement Antisera............................$175.00 USD/Vial Includes 1 vial Antisera (IRM 914) only Sufficient for ASTM D 6499 testing of 7500 samples when used with the proper amount of NR Protein (IRM 913)

Replacement Package NR Protein .........................................$716.00 USD/15 Vials Includes 15 vials NR Protein (IRM 913) only Sufficient for ASTM D 6499 testing of 7500 samples when used with the proper amount of Antisera (IRM 914)

Single Replacement NR Protein .......................................$64.00 USD/Vial Includes 1 vial NR Protein (IRM 913) only Sufficient for ASTM D 6499 testing of 500 samples when used with the proper amount of Antisera (IRM 914)