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Nordic was established in 1964 as a firm for manufacturing and trade of pharmaceutical and chemical compounds. The focus shifted to immunology and the production of antisera became the main activity. Cooperation with the Institutes of the Dutch Health Organisation for Applied Scientific Research (TNO) resulted in many specific antisera to human, mouse, monkey and rat immunoglobulin classes and their subclasses. These are still important items in the product range of Nordic. Of interest may also be a wide range of polyclonal antisera to other serum proteins of different animal species.
Early techniques were based on precipitation reactions of antisera with the antigen. Although underestimated nowadays, immunoelectrophoresis, single and double radial immunodiffusion (Mancini, Ouchterlony) are still valuable techniques. Along with the development of more sensitive techniques Nordic Laboratories produce fluorescent and horseradish peroxidase conjugates as well as biotin conjugates for labels of the customers own choice.
Over the years we have established working reference standards of reactivity, purity and stability, which are permanently reviewed in the light of current progress. Our aim is to provide the customer a wide scale of basic immunological products of high quality, also suited for more traditional techniques and small-scale applications, as well as support based on many years of experience.
Today we offer a range of over 1700 products for fundamental and applied research of human and veterinary systems. It includes a unique series of polyclonal antisera against enzymes as unlabelled IgG fractions, biotin conjugates and affinity purified antibody fractions.
FIX&PERM® Cell Fixation and Permeabilization Kit (Cat# GAS-002)
Intracellular staining of protein biomarkers using antibodies in flow cytometry has opened up a diversity of new options for phenotypic and functional characterisation of cells in both research and clinical diagnosis.
Indeed, definitive phenotypic identification of certain cell types can require labelling of both cell surface and intracellular markers. Additionally, the cytoplasmic localisation of typical membrane molecules can prove to be a most reliable lineage marker, such as cytoplasmic CD3 and CD22 in undifferentiated leukaemia.
In such cases, and for many other cell biological and functional investigations, intracellular immunostaining is a necessity. For this purpose Nordic-MUbio offers a class leading cell fixation and permeabilization kit – FIX&PERM® – which offers a number of advantages over other methods for intracellular staining of cells in flow cytometry.
- Mildly fixes cells, preserving their flow cytometric scatter characteristics
- Allows simultaneous characterisation 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
- Stringent QC procedures – the quality of each lot is determined using well-defined blood samples and subsequent comparison of scatter characteristics of obtained leukocyte populations, ensuring consistent and reliable results lot after lot
- A range of intracellular antibodies with optimised protocols for use with FIX&PERM®
FIX&PERM® is a simple 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 labelled antibodies, allowing staining of intracellular structures such as cytoplasmic or nuclear enzymes, oncoproteins, cytokines, immunoglobulins, etc.
For each sample to be analysed use 50 µl of whole blood, bone marrow or mononuclear cell suspension in a 5 ml tube
- 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 re-suspend cells in sheath fluid for immediate analysis or re-suspend cells in 0.5 ml 1.0 % formaldehyde and store at 2-8°C in the dark
- Analyse fixed cells within 24 hours
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