ADG FIX&PERM^®

Fixation and Permeabilization Kit

From AN DER GRUB, the pioneer in the field • For simultaneous detection of intracellular of intracellular flow cytometry. and surface antigens.

Worldwide recognized as superior reagent • Excellent scatter characteristics. for intracellular flow cytometry.
• Reliable and reproducible results.

Well-suited for leukemia and lymphoma immunophenotyping. • Rapid procedure.
May be used with isolated cells or whole blood. • Easy to use and cost effective.

Intended use

This FIX&PERM^®Cell Permeabilization Kit contains 2 reagents: Fixation Medium (Reagent A) and Permeabilization Medium (Reagent B). It is intended for first fixing cells in suspension with Reagent A and then permeabilizing the cell membranes with Reagent B. This procedure gives antibodies access to intracellular structures and leaves the morphological scatter characteristics of cells intact. Specific formulations reduce background staining and allow simultaneous addition of permeabilization medium and fluorochrome labeled antibodies.

FIX&PERM^®is suitable for the analysis of normal and malignant leukocyte populations derived from various human biological samples (blood, bone marrow and others) using flow cytometry. Results must be put within the context of other diagnostic tests as well as the clinical history of the patient by a certified professional before final interpretation.

Vortex at low speed for 1–2 seconds.
Incubate for 15 minutes at room temperature.
Wash cells with phosphate buffered saline as described above.
Remove supernatant and resuspend cells in sheath fluid for immediate analysis or resuspend cells in 0.5 ml

1.0 % formaldehyde and store them at 2–8°C in the dark. Analyze fixed cells within 24 hours.

Samples

Biological fluids (blood, bone marrow, and others) must be collected under sterile conditions. Anticoagulation with EDTA or heparin is recommended. The samples should be stored at room temperature until used. For optimal results, samples should be processed and analyzed within 24 hours. Samples with high numbers of non-viable cells might cause false results, such cases require determination of cell viability with

e.g. propidium iodide.

results. Lysis of red cells might be impossible for various reasons. In such instances it is recommended to isolate mononuclear cells (MNC) via density gradient centrifugation prior to staining.

Results will be correct and reproducible as long as the procedures used respect the technical recommendations and obey good laboratory practice.

The FIX&PERM^®solutions are provided in a concentration that will allow to fix and permeabilize human hematopoietic cells. It is therefore strongly recommended to stick to the working protocol in terms of concentration and volume regarding cells and antibody.

The properties of FIX&PERM^®have been determined using EDTA anti-coagulated peripheral blood.

Precautions

For professional users only. Reagent A of FIX&PERM^®Cell Permeabilization Kit contains fomaldehyde. Formaldehyde is toxic, allergenic and a suspected carcinogen. Avoid contact with eyes, skin and clothing. Proper handling procedures are recommended.

Storage

FIX&PERM^®Cell Permeabilization Kit reagents should be stored and used at room temperature. Stability of the reagent: Please refer to the expiry date printed onto the vial. The use of the reagent after the expiration date is not recommended. Do not use reagents if a precipitate should form or discoloration occurs.

If unexpected results are obtained which cannot be attributed to differences in laboratory procedures, please contact us.

FIX&PERM^®Cell Permeabilization Reagents

FIX&PERM® Kit	2 x 20 ml	200 Tests	GAS-002	Gerichtsberg 28, 2572 Kaumberg, Austria
FIX&PERM® Sample Kit	2 x 5 ml	50 Tests	GAS-002M	Ortliebgasse 25, 1170 Vienna, Austria
				Phone: +43 1 489 42 14-0
FIX&PERM® Solution A (Fix)	100 ml	1.000 Tests	GAS-002A-1	Fax: +43 1 489 42 14-50
FIX&PERM® Solution B (Perm)	100 ml	1.000 Tests	GAS-002B-1	office@andergrub.com www.andergrub.com

AN DER GRUBs monoclonal Antibodies optimized for use with FIX&PERM^®Cell Permeabilization Kit:

Single IC Antibodies/Conjugates

Single Antibodies/Conjugates for intracellular antigen detection

CD3	Mouse	UCHT1	IgG1	purified	0.2 mg	GM-4011
				FITC	100 Tests	GM-4012
				PE	100 Tests	GM-4013
CD22	Mouse	RFB4	IgG1	purified	0.2 mg	GM-4051
				PE	100 Tests	GM-4053
Lactoferrin	Mouse	4C5	IgG1	purified	0.2 mg	GM-4111
				PE	100 Tests	GM-4113
Myeloperoxidase (MPO-C2)	Mouse	8E6	IgG1	purified	0.2 mg	GM-4191
				FITC	100 Tests	GM-4192
				PE	100 Tests	GM-4193
Lysozyme	Mouse	LZ-2	IgG1	purified	0.2 mg	GM-4131
				FITC	100 Tests	GM-4132
CD68	Mouse	Ki-M7	IgG1	FITC	100 Tests	GM-4152
IL-1 beta	Mouse	FIB3	IgG1	FITC	100 Tests	GM-4162
IFN-gamma	Mouse	GZ4	IgG1	FITC	100 Tests	GM-4172

Errors and omissions exempt.

Combi IC Antibodies/Conjugates

Combi Conjugated for intracellular antigen detection

Negative Control	Mouse		IgG1	FITC	50 Tests	GIC-201
	Mouse		IgG1	PE
Lysozyme	Mouse	LZ-2	IgG1	FITC	50 Tests	GIC-206
Lactoferrin	Mouse	4C5	IgG1	PE
Myeloperoxidase (MPO-C2)	Mouse	8E6	IgG1	FITC	50 Tests	GIC-212
Lactoferrin	Mouse	4C5	IgG1	PE
Myeloperoxidase (MPO-C2)	Mouse	8E6	IgG1	FITC	50 Tests	GIC-213
CD3	Mouse	UCHT1	IgG1	PE
Myeloperoxidase (MPO-C2)	Mouse	8E6	IgG1	FITC	50 Tests	GIC-214
CD22	Mouse	RFB4	IgG1	PE

Distributed by:

ADG FIX&PERM®