Chrono-Log 490 4+4 Instruction Manual page 10

a. Water
Catalog No.: 398
Supplied As: Purified Water 15mL
Sterile distilled, bottled water suitable for CHRONO-
PAR® Reagent preparation.
Pyrogen free (ATP free) for reconstituting reagents
and not containing preservatives/additives such as
benzyl alcohol which inhibits platelet function. Do not
use water from the lab purification system.
b. Saline
Catalog No.: 397
Supplied As: Saline (0.9%) 15mL
Sterile, physiological, saline for CHRONO-PAR® Reagent
preparation.
Avoid blood bank saline because it may be an
incorrect osmolarity. Cell counter diluents are not
suitable because they contain EDTA, which inhibits
platelet aggregation. Infusion salines are
inappropriate because they contain benzyl alcohol (or
other preservatives). Such preservatives/additives
inhibit platelet function.
c. ADP
Catalog No.: 384
Supplied As: 2.5 mg of lyophilized preparation of
adenosine diphosphate.
Stock Conc.: 1 mM
Stock Storage: Store frozen at below 0°C
Stock Shelf life: Until expiration date.
Working Conc.: 1 mM
Working Storage: 2-8°C
Working Shelf life: 8 hours
Reconstitute with 5.0 mL of irrigation grade
physiological saline.
Preparation: Tap vial gently to get contents to the
bottom. Reconstitute with 5 mL of irrigation grade
physiological saline. Allow to sit for 10 minutes with
occasional inversion. Add 5 µL of reagent to 500 µL
sample or 2.5 µL of reagent to 250 µL sample for a final
concentration of 10 µM. Normal aggregation is seen in
PRP with final concentrations of 5-10 µM.
Stability: The reconstituted ADP reagent can be stored
frozen at -70°C volumes suitable for a days testing for
one year or until expiration date, whichever comes
first.
d. Arachidonic Acid
Catalog No.: 390
Supplied As: Minimum of 10 mg of Arachidonic Acid
with a purity of better than 99%. Albumin contains
100mg of bovine albumin, fraction V powder, 96 to 99%
pure.
Stock Conc.: 50 mM
Document # 49044IM1
Revision 7.5
Dated February 16, 2017
Stock Storage: Frozen below -20°C for Arachidonic Acid,
Refrigerate at 2-8°C for albumin.
Stock Shelf Life: Until expiration date.
Working Conc.: 50 mM
Working Storage: 2-8°C [in the dark]
Working Shelf Life: 8 hours
Reconstitute with 0.7 mL of the saline-albumin
solution.
Preparation: First tap contents gently to the bottom of
the vial of albumin. Reconstitute the albumin with 1
mL of irrigation grade physiological saline. Allow to sit,
then mix with occasional swirling. Allow 15 to 30
minutes for the albumin to fully absorb the saline
(check visually). The Arachidonic Acid in the vial is an
oily drop which must be shaken or tapped to the
bottom of the vial. Break vial tip with Cap Cracker
supplied. Pipette reconstituted albumin into both the
tip and body of the vial in 100 µL aliquots to a total
volume of 700 µL. Make sure any Arachidonic Acid
remaining on the tip or body of the vial is mixed by
rotating the vial as the albumin is added. Repeat a few
times in each section of the vial then vigorously mix
the Arachidonic Acid into the albumin using a transfer
pipette. Combine the suspension from the tip with
that in the body of the vial and continue mixing until
the Arachidonic Acid suspension reaches maximum
turbidity. Transfer reagent to micro centrifuge tube
and vortex for 5 minutes. The reconstituted
Arachidonic Acid suspension should appear very milky
with numerous small bubbles.
Add 5 µL of reagent to 500 µL PRP sample or 2.5 µL of
reagent to 250 µL sample for a concentration of 0.5
mM. Normal aggregation is seen with final
concentrations of 0.5 -1.0 mM.
Stability: The reconstituted Arachidonic Acid can be
stored frozen at -70°C in the dark in 100 µL volumes
for 3 months or until expiration date, whichever comes
first. When stored frozen at -20°C in the dark
Arachidonic Acid is stable for 1 month or until
expiration date, whichever comes first. Aliquots can be
hand thawed and vigorously re-suspended with a
vortex mixer just before use.
e. Collagen
Catalog no.: 385
Supplied as: 1 mg of native Collagen fibrils (type I) from
equine tendons suspended in isotonic glucose solution
of ph 2.7/vial.
Stock Conc.: 1 mg/mL
Stock Storage: Refrigerate at 2-8°C
Stock Shelf Life: Until expiration date.
Working Conc.: 1 mg/mL
Working Storage: 2-8°C
Working Shelf life: Until expiration date.
Preparation: Collagen can be used directly as supplied.
Invert or swirl vial before use, as Collagen fibrils are in
suspension. Do not freeze. If required, Collagen can
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