Intracellular staining of protein biomarkers using antibodies in flow cytometry has opened diverse new pathways for phenotypic and functional characterization of cells in both research and clinical diagnosis.
Definitive phenotypic identification of certain cell types can require labeling of both cell surface and intracellular markers. Additionally, the cytoplasmic localization of typical membrane molecules can prove to be a most reliable lineage marker, such as cytoplasmic CD3 and CD22 in undifferentiated leukemia.
In such cases, and for many other cell biological and functional investigations, intracellular immunostaining is a necessity. For this purpose, we offer an industry-leading cell fixation and permeabilization kit – FIX&PERM® – which offers several advantages over other methods for intracellular staining of cells in flow cytometry.
FIX&PERM® is a simple flow cytometry cell preparation procedure making use of two reagents. Reagent A gently fixes cells, while Reagent B permeabilizes them. The specific formulations reduce background and allow simultaneous addition of permeabilization medium and fluorochrome-labeled antibodies, allowing staining of extracellular surface markers and intracellular structures such as cytoplasmic enzymes, nuclear proteins, oncoproteins, cytokines and immunoglobulins.
- Mildly fixes cells, preserving their flow cytometric scatter characteristics
- Allows simultaneous characterization of both intracellular and cell surface markers
- Rapid technique – whole procedure can be carried out in less than one hour, ready for immediate analysis or storage for 24 hours
- Proven technology – initially characterized in Knapp et al. in 1994, independently validated in studies such as Groeneveld et al. in 1996 and Lanza et al. in 1997, and cited in hundreds of peer-reviewed publications since
- Stringent QC procedures – the quality of each lot is confirmed using well-defined blood samples and subsequent comparison of scatter characteristics and immunostaining patterns, ensuring consistent and reliable results each time
- Stable – our formulations do not contain volatile organic solvents, enabling storage at ambient temperature during shipment and in laboratories
- Flexible – can be used with most monoclonal antibody conjugates, as well as our range of antibodies with optimized protocols for use with FIX&PERM®
- CE/IVD registered kits available for European customers in a clinical laboratory setting (contact us for further information)
|GAS-002||FIX&PERM® Cell Fixation and Permeabilization Kit||Buffers and Reagents|
|GAS-002-1||FIX&PERM® Kit 1000||Buffers and Reagents|
|GAS-002A-1||FIX&PERM® Solution A (Fix)||Buffers and Reagents|
|GAS-002B-1||FIX&PERM® Solution B (Perm)||Buffers and Reagents|
|GAS-002M||FIX&PERM® Sample Kit||Buffers and Reagents|
|GAS-002A-1-CE/IVD||FIX&PERM® Solution A (Fix) (CE/IVD)||Buffers and Reagents|
|GAS-002-1-CE/IVD||FIX&PERM® Kit 1000 Cell Fixation and Permeabilization Kit (CE/IVD)||Buffers and Reagents|
|GAS-002B-1-CE/IVD||FIX&PERM® Solution B (Perm) (CE/IVD)||Buffers and Reagents|
|GAS-002-CE/IVD||FIX&PERM® Cell Fixation and Permeabilization Kit (CE/IVD)||Buffers and Reagents|
|GAS-002M-CE/IVD||FIX&PERM® Sample Kit (CE/IVD)||Buffers and Reagents|
If you are a distributor/OEM interested in FIX&PERM®, please contact us.
- Add 100 µl of Reagent A (Fixation Medium, stored and used at room temperature)
- Incubate for 15 minutes at room temperature
- Add 5 ml 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 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 at 2-8°C in the dark
- Analyze fixed cells within 24 hours
|Negative control||Vl-AP||IgG1||FITC-labeled||50 tests||GIC-201|
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