ASTM Committee D11 - Rubber and Rubber Products 'Document Sheet' for Industry Reference Material IRM-914 Anti-AL Antisera

Introduction
Industry Reference Material (IRM) 914 is the designation for the ammoniate natural rubber (NR) protein reference antisera that is used as a reference antisera according to the procedures of D6499. Details on the evaluation of this material are found in Research Report D11-1094.

Properties
Partially purified ammoniated natural rubber proteins (ASTM natural rubber (NR) standard antigen) were used as the antigens for eliciting the immune response in rabbits. Rabbits were immunized on day 1 and given booster injections on weeks 4, 5, and 7.

Property ASTM Designation Limits/Targets

Animal	D-6499	Rabbits
Antigen	D-6499	Ammoniated NR reference protein
Adjuvant	D-6499	Freund's complete and incomplete
Titer	D-6499	Minimum 1/8,300
Western Blot Profile	D-6499	Reactivity with entire spectrum of SDS-PAGE visible AL protein
Inhibition ELISA reactivity	D-6499	Comparative analysis of extracts from a minimum of 5 NR products

Accepted Reference Value (AR Value)
This reagent is used in the D6499 procedure and the assigned protein value is determined by comparison to the AL protein reference material. The assigned reference value is given as a item (1/15,000), determined by one laboratory. The performance of the antisera at 1/15,000 was tested by seven laboratories analyzing a 4 ug/ml sample of AL on 2 separate plates (mean % inhibition of duplicate determinations).

(1) AR-Value in Percent Inhibition AR-Value (%) +/- 2 Sigma Min Max

Range

83.6	3.5	80.1	87.1	7.0
76.3	3.5	72.8	79.8	7.0
68.0	3.5	64.5	71.5	7.0

Specific Conditions Under Which the IRM 914 is to be Stored
The material is stored as 1 ml aliquots at -20º C in 20% glycerol as a cryo-preservative. The vials must be stored at -20ºC and shipped by courier (overnight) on dry ice. The shelf life of the antisera has not been determined, but should have a minimum shelf life of 10 years provided the -20ºC conditions are maintained. To ensure quality the sera should be tested for its titer every 6 months to ensure that significant degradation has not occurred. The extent of degradation should not be greater than the typical error found in between laboratory testing (ASTM D6499).

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

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

Replacement Antisera
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).
$175.00 USD/Vial

Replacement Package NR Protein
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).
$716.00 USD/15 Vials

Single Replacement NR Protein
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).
$64.00 USD/Vial

Domestic shipments can be made within 48 hours of receipt of purchase order. International shipments can be made within 48 hours of receipt of funds in US currency. Wire transfer, Mastercard/VISA also accepted.