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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 
ReaFix Fixative Solution used   for   the   fixation   of   cell   preparations   before 
10X Concentrate analysis by flow cytometry.

Catalog No. 25325­ 00 Note  ­  Follow   appropriate   instructions   as 


given   in   the   respective   product   inserts   for 
1.  PRODUCT DESCRIPTION information   on   instruments,   sample   collection   and 
sample processing.
The   ReaFix   fixative   solution   is   a   10X 
concentrated formaldehyde based fixative solution in  4.1 Dilution Instructions: Preparation of the 
Phosphate   buffer   saline   (PBS).   It   is   intended   for  1X Working Solution
fixing   cell   preparations   to   enable   a   fix­no   wash 
protocol   in   applications   that   involve  Dilute   the   10X   concentrate   1:10   deionized 
immunofluorescent   staining   and   analysis   by   flow  water at room temperature (20° to 25°C) to prepare 
cytometry. 1X working fixative solution according to Table 1. 
Mix well.
2.  INTENDED USE  Prepare   an   adequate   amount   of   the   1X 
working   solution   depending   on   the   reagent   and 
The ReaFix fixative solution is recommended  volume to be used per tube.
for use in fixation of cell preparations stained with 
ReaMetrix   reagents   such   as   Rea   T­Count   reagent  Table 1: Dilution Instructions for Preparation of 1X  
(Dual Tube Catalog Number: 25124­00; Single Tube  Working Fixative solution
Catalog   Number:   25242­00)   and   ReaPan   Thrombo 
reagent (Catalog Number: 25232­00) which are to be  Required Volume 
analyzed by flow cytometry.  of  1X Working  5 10 15 20 25
Use   of   ReaFix   fixative   solution   in   the 
solution (mL)
processing   steps   for   analysis   of   the   sample 
Volume of 10X 
preparations   to   be   analyzed   using   the   above­
Concentrate 
mentioned   reagents   (Section   1)   is   highly  0.5 1.0 1.5 2.0 2.5
ReaFix Fixative 
recommended. This ensures the sufficient fixing of 
the   immunofluorescent   stained   populations   without  Solution (mL)
Volume of 
lysing the red blood cells 
Deionized Water  4.5 9.0 13.5 18.0 22.5
3.  PRINCIPLE  (mL)

When   whole   blood   is   added   to   the 


monoclonal   antibody   reagent,   the   fluorochrome­ 4.2 Storage Condition and Stability
labeled antibodies in the reagent bind specifically to 
The 1X working fixative solution should be prepared 
target surface antigens. The stained samples are then 
daily;   the   unused   1X   Fixative   solution   should   be 
treated with ReaFix fixative solution which stabilizes 
discarded at the end of the day. Do not refrigerate.
cell proteins by binding covalently between the free 
amine groups, without lysing the red blood cells. The 
4.3 Evidence of Deterioration
stained fixed cell preparations are suitable for flow 
cytometric analysis. The normal appearance of ReaFix Fixative Solution 
reagent is that of a clear liquid. Any change in the 
4.  REAGENT  physical appearance of the reagent, major variation in 
values for control subjects may indicate deterioration,  CD45   gating   for   persons   infected   with 
and the reagent should not be used. human immunodeficiency virus. Centers for 
Disease   Control   and   Prevention.   MMWR 
Recomm Rep 2003 Jan 31; 52(RR­2):1­13.

5.  PRECAUTIONS 2. Mandy FF, Brando B. Enumeration of 
Absolute Cell Counts Using 
1. The   ReaFix   Fixative   Solution   contains 
Immunophenotypic Techniques. Current 
Formaldehyde.   Formaldehyde   is   a   potential 
Protocols in Cytometry (2000) 6.8.1­6.8.26.
carcinogen   and   causes   irritation   to   the   eyes   and 
skin. Avoid inhalation, contact with eyes, skin and 
3. Bossuyt   X,  Marti   GE,  Fleisher   TA: 
clothing. Wear suitable protective clothing. Never 
Comparative   analysis   of   whole   blood   lysis 
pipette   by   mouth;   if   swallowed,   seek   medical 
methods   for   flow   cytometry.   Cytometry 
attention immediately.
30:124–133, 1997.
2. All   blood   specimens   are   considered   biohazards. 
Handle them as if they are capable of transmitting  4. Dressler, L.G., "Specimen handling, storage, 
infection and dispose off with proper precautions  and   preparation",   1997,   Curr.   Protocols 
in accordance with governmental regulations. Cytometry, Chapter 5, 5.0.1­ 5.2.15.

3. The addition of  precise volume of deionized 
7.  WARRANTY
water is critical to obtain correct concentration of 
the 1X working solution. Use a calibrated pipette  The product is warranted only to conform to 
and   operate   according   to   the   manufacturer’s  the quantity and contents stated on the label at the 
instructions. time   of   delivery   to   the   customer.   There   are   no 
4. The use of temperatures, incubation and vortexing  warranties, expressed or implied, that extend beyond 
times   other   than   those   specified   in   the  the   description   on   the   label   of   the   product. 
corresponding product inserts may give erroneous  ReaMetrix’ sole liability is limited to replacement of 
results. the   products.   ReaMetrix   is   not   liable   for   property 
damage, personal injury, or economic loss caused by 
5. The  ReaFix   Fixative   Solution  contains 
the product.
Formaldehyde   which   is   highly   volatile.   Do   not 
leave the lid in a loose or open position. The lid 
CUSTOMER SUPPORT INFORMATION
should be tightly sealed after use.
6. Avoid microbial contamination of the reagent or  ReaMetrix India Pvt. Ltd.
erroneous results may occur. 50­B, II Phase, Peenya Industrial Area
Peenya, Bangalore­560058, India
7. Results   obtained   with   flow   cytometry   may   be 
Ph: 91­80­28378693/5,
erroneous   if   the   instrument   is   misaligned, 
Fax: 91­80­41172451
compensation   spillovers   are   not   correctly 
E­mail: info@reametrix.com
compensated or the gate is improperly set.
www.reametrix.com
8. Reproducible results will be obtained as long as 
Rev No. 1.0, 27­Apr­09
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   single­platform 
absolute   CD4+   T­cell   determinations   with