Bio-Rad Human MMP and TIMP Assays - Manual (Page 15)

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2. Add 150 µl of the appropriate diluent to tubes S2–S8 (Figure 3).
3. Vortex reconstituted standards at medium speed for 5 sec before
removing any volume. Transfer 75 µl to the S2 tube containing the
chosen diluent. Vortex for 5 sec.
4. Use a new pipet tip to transfer 75 µl from the S2 tube to the S3 tube.
Vortex for 5 sec.
5. Continue with 1:3 (threefold) serial dilutions as shown in Figure 3.
6. Use reconstituted and diluted standards and controls immediately.
Do not freeze for future use.
Fig. 3. Preparing a threefold dilution series with a single reconstituted standard.
Note: For samples with very low endogenous analytes, prepare one additional standard point
(S9) to extend the bottom end of the standard curve.
6. Prepare Samples
General guidelines for preparing different sample types are provided
here. For more information, consult publications listed in Bio-Rad bulletin
#5297, available for download at www.bio-rad.com, or contact Bio-Rad
Technical Support.
n
Once thawed, keep samples on ice. Prepare dilutions just prior to the
start of the assay and equilibrate to room temperature before use
n
Prepare sample dilutions in microcentrifuge tubes. Alternatively, if a
multichannel pipet will be used to load the plate, aliquot the required
volumes into Titertube
® micro test tubes
n
Do not freeze diluted samples
150 150 150 150 150 150 150 150
S1 S2 S3 S4 S5 S6 S7 S8 Blank
Diluent, µl
75 75 75 75 75 75 75 Transfer Volume, µl
Reconstituted
Standard