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

8/14/2019 PI 25325 00 ReaFix Fixative Solution

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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: info@reametrix.com

www.reametrix.com

Rev No. 1.0, 27-Apr-09

http://www.reametrix.com/http://www.reametrix.com/