Download - PI 25325 00 ReaFix Fixative Solution
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8/14/2019 PI 25325 00 ReaFix Fixative Solution
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ReaFix Fixative Solution
10X ConcentrateCatalog No. 25325- 00
1. PRODUCT DESCRIPTION
The ReaFix fixative solution is a 10X
concentrated formaldehyde based fixative solution in
Phosphate buffer saline (PBS). It is intended for
fixing cell preparations to enable a fix-no wash
protocol in applications that involve
immunofluorescent staining and analysis by flow
cytometry.
2. INTENDED USE
The ReaFix fixative solution is recommended
for use in fixation of cell preparations stained with
ReaMetrix reagents such as Rea T-Count reagent
(Dual Tube Catalog Number: 25124-00; Single Tube
Catalog Number: 25242-00) and ReaPan Thrombo
reagent (Catalog Number: 25232-00) which are to be
analyzed by flow cytometry.
Use of ReaFix fixative solution in the
processing steps for analysis of the sample
preparations to be analyzed using the above-
mentioned reagents (Section 1) is highly
recommended. This ensures the sufficient fixing of
the immunofluorescent stained populations without
lysing the red blood cells
3. PRINCIPLE
When whole blood is added to the
monoclonal antibody reagent, the fluorochrome-
labeled antibodies in the reagent bind specifically to
target surface antigens. The stained samples are then
treated with ReaFix fixative solution which stabilizes
cell proteins by binding covalently between the free
amine groups, without lysing the red blood cells. The
stained fixed cell preparations are suitable for flowcytometric analysis.
4. REAGENT
The ReaFix fixative solution is formulated as
a buffered solution and contains Formaldehyde as a
fixative agent. It is provided as a 10X concentrate. A
working solution (1X) of this concentrate is to be
used for the fixation of cell preparations before
analysis by flow cytometry.
Note - Follow appropriate instructions as
given in the respective product inserts for
information on instruments, sample collection and
sample processing.
4.1 Dilution Instructions: Preparation of the
1X Working Solution
Dilute the 10X concentrate 1:10 deionized
water at room temperature (20 to 25C) to prepare
1X working fixative solution according to Table 1.
Mix well.
Prepare an adequate amount of the 1X
working solution depending on the reagent and
volume to be used per tube.
Table 1: Dilution Instructions for Preparation of 1X
Working Fixative solution
Required Volume
of 1X Working
solution (mL)
5 10 15 20 25
Volume of 10X
Concentrate
ReaFix Fixative
Solution (mL)
0.5 1.0 1.5 2.0 2.5
Volume of
Deionized Water
(mL)
4.5 9.0 13.5 18.0 22.5
4.2 Storage Condition and Stability
The 1X working fixative solution should be prepared
daily; the unused 1X Fixative solution should be
discarded at the end of the day. Do not refrigerate.
4.3 Evidence of Deterioration
The normal appearance of ReaFix Fixative Solution
reagent is that of a clear liquid. Any change in the
physical appearance of the reagent, major variation in
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values for control subjects may indicate deterioration,
and the reagent should not be used.
5. PRECAUTIONS
1. The ReaFix Fixative Solution containsFormaldehyde. Formaldehyde is a potential
carcinogen and causes irritation to the eyes and
skin. Avoid inhalation, contact with eyes, skin and
clothing. Wear suitable protective clothing. Never
pipette by mouth; if swallowed, seek medical
attention immediately.
2. All blood specimens are considered biohazards.
Handle them as if they are capable of transmitting
infection and dispose off with proper precautionsin accordance with governmental regulations.
3. The addition of precise volume of deionized
water is critical to obtain correct concentration of
the 1X working solution. Use a calibrated pipette
and operate according to the manufacturers
instructions.
4. The use of temperatures, incubation and vortexing
times other than those specified in the
corresponding product inserts may give erroneousresults.
5. The ReaFix Fixative Solution contains
Formaldehyde which is highly volatile. Do not
leave the lid in a loose or open position. The lid
should be tightly sealed after use.
6. Avoid microbial contamination of the reagent or
erroneous results may occur.
7. Results obtained with flow cytometry may be
erroneous if the instrument is misaligned,compensation spillovers are not correctly
compensated or the gate is improperly set.
8. Reproducible results will be obtained as long as
the procedure used is in accordance with this
package insert and with good laboratory practice
guidelines.
6. REFERENCES
1. Mandy FF, Nicholson JK, McDougal JS.
Guidelines for performing single-platform
absolute CD4+ T-cell determinations with
CD45 gating for persons infected with
human immunodeficiency virus. Centers for
Disease Control and Prevention. MMWR
Recomm Rep 2003 Jan 31; 52(RR-2):1-13.
2. Mandy FF, Brando B. Enumeration of
Absolute Cell Counts Using
Immunophenotypic Techniques. Current
Protocols in Cytometry (2000) 6.8.1-6.8.26.
3. Bossuyt X, Marti GE, Fleisher TA:
Comparative analysis of whole blood lysis
methods for flow cytometry. Cytometry
30:124133, 1997.
4. Dressler, L.G., "Specimen handling, storage,
and preparation", 1997, Curr. Protocols
Cytometry, Chapter 5, 5.0.1- 5.2.15.
7. WARRANTY
The product is warranted only to conform to
the quantity and contents stated on the label at the
time of delivery to the customer. There are no
warranties, expressed or implied, that extend beyond
the description on the label of the product.
ReaMetrix sole liability is limited to replacement of
the products. ReaMetrix is not liable for property
damage, personal injury, or economic loss caused by
the product.
CUSTOMER SUPPORT INFORMATION
ReaMetrix India Pvt. Ltd.
50-B, II Phase, Peenya Industrial Area
Peenya, Bangalore-560058, India
Ph: 91-80-28378693/5,
Fax: 91-80-41172451
E-mail: [email protected]
www.reametrix.com
Rev No. 1.0, 27-Apr-09
http://www.reametrix.com/http://www.reametrix.com/