CD4 easy count kit

Product Name CD4 easy count kit IVD

Code No. 05-8401

Content Packing contains reagents for 100 tests:

• 100 ml no lyse buffer
• 2000 µl CD4 mAb PE (MEM-241, PE-conjugated
monoclonal antibody to human CD4)

General Information: Patients with AIDS exhibit T-cell
lymphopenia, a loss of CD4+ lymphocytes and a relative
increase in the CD8+ subtype and in the CD3+CD4-CD8-
subtype. A precise enumeration of CD4+ T-cells is
necessary for a reliable and controlled antiretroviral therapy
and patient monitoring.
Specificity and Antigen Distribution: The mouse
monoclonal antibody MEM-241 recognizes the human CD4
antigen, a transmembrane glycoprotein (55 kDa) of the
immunoglobulin supergene family, present on a subset of
T-lymphocytes (“helper/inducer” T-cells) and also
expressed at a lower level on monocytes, tissue
macrophages and granulocytes. Approximately 20-60% of
human peripheral blood mononuclear cells as well as a
subpopulation of monocytes but with a weaker signal are
stained. The antibody has been studied at the 8th
International Workshop on Human Cell Differentiation
Molecules HCDM (former HLDA VIII), May 2006, Québec,
Canada. CD4 is the primary cellular receptor for the human
immunodeficiency virus (HIV).
Clone MEM-241
Isotype Mouse monoclonal IgG1
Applications:
• Identification and enumeration of the CD4+
helper/inducer T-lymphocytes subset
• Studies of HIV infection. Patients infected with HIV loose
CD4+ helper/inducer lymphocytes
• Characterization of subtypes of T-cell leukemias and
lymphomas
• For in vitro diagnostic use
Method:
• Add 20 µl whole blood (EDTA as anticoagulant) to a
Partec test tube
• Add 20 µl of CD4 mAb PE. Mix gently and incubate for
15 minutes at room temperature protected from light
• Add 800 µl of no lyse buffer and shake or vortex gently
• Analyze blood sample on a Partec device
Flow Cytometric Analysis: CD4-PE fluorescence can be
analysed on a Partec Flow Cytometer with an excitation light
source of 488 nm or 532 nm (blue or green solid state laser).
To count CD4+ T-cells transfer the test tube with 840 µl of the
ready prepared blood sample (see Method) to your Partec
Flow Cytometer and carry out a count analysis. For detailed
information on the handling and calibration of the instrument
please refer to the operating manuals and application notes.
The blood preparation allows one count procedure on the
same sample. Following the above procedure the dilution
factor is 42. In the CyFlow® Counter and CyFlow® SL_3 this
dilution factor is already set or can be directly set by the
instrument operator in the instrument software. Counting
results will be displayed automatically as CD4+ T-cells per µl
whole blood.
Recommended and Required Materials:
• Partec Flow Cytometry instrument (e.g. CyFlow® Counter
or CyFlow® SL_3)
• Partec test tubes (Code No. 04-2000)
• Micropipettes and pipette tips (e.g. Eppendorf, Code No.
3112 000.029 and 3111 000.0165)
• Powder-free latex gloves (e.g. Safeskin, Code No. 545-950-
06)
• Venous Blood Collection System with EDTA as
anticoagulant (e.g. Greiner Bio-One: Vacuette® EDTA
Tubes, K3E/EDTA K3, 3 ml, Code No. 454217, Vacuette®
Blood Collection Needles 38 x 0.8 mm, Code No. 450076,
Vacuette® Tube Holders, Code No. 450201)
Handling and Storage: CD4 mAb PE is supplied in 2 ml of
phosphate-buffered saline (PBS) pH 7.4, containing BSA and
0.09% sodium azide, sufficient for 100 tests. 20 µl of CD4
mAb PE are sufficient for labelling 1x106 cells. When stored at
2-8°C in the dark, the CD4 easy count kit is stable until the
expiration date printed on the kit label. Do not freeze or
expose to elevated temperatures.

For in vitro diagnostic use with Partec IVD Flow Cytometers.

Contact Information:

Partec GmbH • Am Flugplatz 13 • D-02828 Görlitz • Germany

Tel +49 3581 8746 0 • Fax +49 3581 8746 70 • E-mail: mail@partec.com
Rev. 010 Date 2010-03-10 05-8401 CD4 easy count kit
 CD4 easy count kit – Page 2

Example of a direct Literature:

counting result using Cassens, U. et al.: Simplified volumetric flow cytometry allows
the CD4 easy count kit - feasible and accurate determination of CD4 T lymphocytes in
printout from a Partec immunodeficient patients worldwide”, Antiviral Therapy 9:
CyFlow® Counter As 395-405 (2004)
depicted in the histogram,
the CD4+ T-cells Göhde, W. et al.: A New Protocol to Follow-up Immune
(prominent peak on the Impairment in HIV/AIDS-Children, Scientific Presentation -
right) can be clearly Abstract No. MoPeLB151C02 at the 3rd IAS Conference on
separated from the CD4+ HIV Pathogenesis and Treatment in Rio de Janeiro, July 2005
monocytes (left peak of Greve, B. et al.: ”A New No-Lyse, No-Wash Flow-Cytometric
weaker fluorescence Method for the Determination of CD4 T Cells in Blood
intensity). The absolute Samples”, Transfus Med Hemoth (2003), 30: 8-13
concentration of CD4+ T-
cells is displayed directly Imade, G. et al.: “Comparison of a New, Affordable Flow
as number of cells/µl Cytometric Method and the Manual Magnetic Bead
whole blood. Technique for CD4 T-Lymphocyte Counting in a Northern
Nigerian Setting”, Clinical and Diagnostic Laboratory
Immunology, Jan. 2005, p. 224-227
Example of a direct CD4 Dieye, T.N. et al.: Absolute CD4 T-Cell Counting in Resource-
counting result using Poor Settings: Direct Volumetric Measurements Versus Bead-
the CD4 easy count kit – Based Clinical Flow Cytometry Instruments, Journal of
display of a Partec Acquired Immune Deficiency Syndromes, Volume 39,
CyFlow® SL_3. The FL2 Number 1, May 1 2005
CD4-PE histogram shows
the precise separation of Fryland, M. et al.: The PARTEC CyFlow counter for CD4+ T-
the CD4+ T-cells (cell cell counting produces high quality results and is robust when
population in the RN1 evaluated under routine field conditions in Malawi, Scientific
range) from the CD4+ Oral Presentation - Abstract No. TuOrB1149 at the XV.
weak monocytes. International AIDS Conference in Bangkok, July 2004

Warnings and Precautions: Reagents contain 0.09%

sodium azide that under acidic conditions yields hydrazoic
acid, an explosive compound. Sodium azide may react with
lead and copper plumbing to form potentially explosive metal
azides. When disposing of sodium azide, flush plumbing with
a large volume of water to prevent build-up. Very toxic if
swallowed. After contact with skin, wash immediately with
plenty of water.
R22: Harmful if swallowed, R32: Contact with acids liberates
very toxic gas, S2: Keep out of reach of children, S13: Keep
away from food, drink and animal feeding stuffs, S36: Wear
suitable protective clothing, S46: If swallowed, seek medical
advice immediately and show this container or label

Handle all specimens and materials as potentially infectious

materials. Decontaminate with 0.05% sodium hypochlorite.
The product should be handled by trained staff only.
Reagents are not sterile. Not for in vivo use. For in vitro use
only.

For in vitro diagnostic use with Partec IVD Flow Cytometers.

Contact Information:

Partec GmbH • Am Flugplatz 13 • D-02828 Görlitz • Germany

Tel +49 3581 8746 0 • Fax +49 3581 8746 70 • E-mail: mail@partec.com
Rev. 010 Date 2010-03-10 05-8401 CD4 easy count kit