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FIX&PERM® Cell Fixation and Permeabilization Kit (CE)


Our Price: $269.00

Product Code: GAS-002

8 x 5ml

Product Details
Product Code: GAS-002
Product: FIX&PERM® Cell Fixation and Permeabilization Kit (CE)
Description: This FIX&PERM® Cell Fixation and 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.

Format: 4 x 5ml vial of Fixation Medium (Reagent A) and 4 x 5ml vial Permeabilization Medium (Reagent B). FIX&PERM® Reagents are designed for use with all commercially available flow cytometers. Alignment and compensation should be performed according to manufacturer´s instructions.

The quality of each FIX&PERM® Lot is determined by fixation and permeabilization of well defined blood samples from representative donors and subsequent comparison of forward and side scatter characteristics of obtained leukocytes, as well as immunostaining efficiency for several membranous and cytoplasmic antigens
Applications: 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. All biological samples have to be handled with caution. Always consider them as potentially infective. Use appropriate precautions such as gloves, lab-coat, etc.

Fixation, permeabilization and staining procedure:

• For each sample to be analyzed add 50 µl of whole blood, bone marrow or mononuclear cell suspension in a 5ml tube • Add 100 µl of Reagent A (Fixation Medium, stored and used at room temperature)
• Incubate for 15 minutes at room temperature
• Add 5ml phosphate buffered saline and centrifuge cells for 5 minutes at 300 g
• Remove supernatant and add to cell pellet 100 µl Reagent B (Permeabilization Medium) and 20 µl of the appropriate Nordic-MUbio monoclonal antibody conjugate
• 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.
Limitations: Flow cytometry should be performed by professional users only. Improper alignment of the flow cytometer, inaccurate compensation of fluorescence leaking into other channels as well as incorrect positioning of regions may lead to false 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.
Storage: FIX&PERM® Cell Fixation and Permeabilization Kit reagents should be stored and used at room temperature. Do not freeze. 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. If reagents are stored under any conditions other than those specified, the conditions must be verified by the user. 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.
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This product is sold for laboratory research use or further manufacturing only and should not be used for human therapeutic or diagnostic applications. The information presented is believed to be accurate; however, said information and products are offered without warranty or guarantee since the ultimate conditions of use and the variability of the materials treated are beyond our control. Nothing disclosed herein is to be construed as a recommendation to use our products in violation of any patents. Under no circumstances shall ARP American Research Products, Inc. be liable for damages, whether consequential, compensatory, incidental or special, strict liability or negligence, breach of warranty or any other theory arising out of the use of the products available from ARP American Research Products, Inc. Nothing contained herein warrants that the use of the products will not infringe on the claims of any patents covering the product itself or the use thereof in combination with other products or in the operation of any process.

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