Download Human Tissue Factor Activity Kit

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Urine: Collect urine using sample pot. Centrifuge samples at 800 x g for
10 minutes and assay. Store samples at -20°C or below for up to 3
months. Avoid repeated freeze-thaw cycles.
Cell Culture Lysates: The cultured cells are lysed and solubilized with 15
mM octyl--D-glucopyranoside at 37°C for 15 minutes. Collect fresh cell
lysates and assay. The samples can be stored at -20°C or below for up to
3 months.
Tissue: Extract tissue samples using 50 mM Tris-buffered saline (pH 8.0)
with 1% Triton X-100 and centrifuge at 14000 x g for 20 minutes. Collect
the supernatant and measure the protein concentration. Dilute the
tissue extract 1:4 into Sample Diluent and assay. Freeze the remaining
extract at -20°C.
Refer to Sample Dilution Guidelines below for further instruction.
Guidelines for Dilutions of 1:100 or Greater
(for reference only; please follow the insert for specific dilution suggested)
1:100
1:10000
A)
4 ul sample: 396 µl buffer(100x)
= 100 fold dilution
A)
B)
Assuming the needed volume is less than
or equal to 400 µl.
Assuming the needed volume is less than
or equal to 400 µl.
1:1000
A)
B)
4 µl sample : 396 µl buffer (100x)
24 µl of A : 216 µl buffer (10x)
= 1000 fold dilution
Assuming the needed volume is less than
or equal to 240 µl.
4 µl sample : 396 µl buffer (100x)
4 µl of A : 396 µl buffer (100x)
= 10000 fold dilution
1:100000
A)
B)
C)
4 µl sample : 396 µl buffer (100x)
4 µl of A : 396 µl buffer (100x)
24 µl of B : 216 µl buffer (10x)
= 100000 fold dilution
Assuming the needed volume is less than
or equal to 240 µl.
Reagent Preparation
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Standard Curve: Reconstitute the rhTF Standard (1500 pM) with 3 ml of
reagent grade water to generate a 500 pM standard stock solution.
Allow the standard to sit for 10 minutes with gentle agitation prior to
making dilutions. Prepare duplicate or triplicate standard points by
serially diluting the standard stock solution (500 pM) 1:2 with Sample
Diluent to produce 250, 125, 62.5, 31.25, 15.63, and 7.81 pM solutions.
Sample Diluent serves as the zero standard (0 pM). Any remaining
solution should be frozen at -20°C and used within 30 days.
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