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. 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 offer a leading cell fixation and permeabilisation kit – The FIX&PERM® – which offers a number of advantages over other methods for intracellular staining of cells in flow cytometry:
FIX&PERM® Key Features
- 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 consisting of two reagents. Reagent A gently fixes cells, while Reagent B permeabilises them. The specific formulations reduce background and allow simultaneous addition of permeabilisation medium and fluorochrome labelled antibodies, allowing staining of intracellular structures such as cytoplasmic or nuclear enzymes, oncoproteins, cytokines and immunoglobulins.
Fix and Perm Reagents Available:
FIX&PERM Sample Kit (CE), (2 x 5ml)
FIX&PERM Kit (CE), (8 x 5ml)
FIX&PERM Kit 1000 (CE), (2 x 100ml)
FIX&PERM Solution A (Fix) (CE), (1 x 100ml)
FIX&PERM Solution B (Perm) (CE), (1 x 100ml)
Products optimised for use with FIX&PERM®
The majority of available monoclonal antibody conjugates can be used with FIX&PERM®. However, in some cases the fixation step may have to be optimised.
Nordic-MUbio offers a number of mouse anti-human conjugated (PE/FITC) monoclonal antibodies optimised for use with FIX&PERM®:
| Antigen | Clone | Isotype | Format | Quantity | Cat. No. |
| CD3 | UCHT1 | IgG1 | Purified | 0.2 mg | GM-4011 |
| FITC-labeled | 100 tests | GM-4012 | |||
| PE-labeled | 100 tests | GM-4013 | |||
| CD22 | RFB4 | IgG1 | Purified | 0.2 mg | GM-4051 |
| FITC-labeled | 100 tests | GM-4052 | |||
| PE-labeled | 100 tests | GM-4053 | |||
| Lactoferrin | 4C5 | IgG1 | Purified | 0.2 mg | GM-4111 |
| PE-labeled | 100 tests | GM-4113 | |||
| Myeloperoxidase | 8E6 | IgG1 | Purified | 0.2 mg | GM-4191 |
| (MPO-C2) | FITC-labeled | 100 tests | GM-4192 | ||
| PE-labeled | 100 tests | GM-4193 | |||
| Lysozyme | LZ-2 | IgG1 | Purified | 0.2 mg | GM-4131 |
| FITC-labeled | 100 tests | GM-4132 | |||
| CD68 | Ki-M7 | IgG1 | FITC-labeled | 100 tests | GM-4152 |
Furthermore, a number of Combi-panels are offered for dual immunostaining protocols:
| Antigen | Clone | Isotype | Format | Quantity | Product Code |
| Negative control | Vl-AP | IgG1 | FITC-labeled | 50 tests | GIC-201 |
| Vl-AP | IgG1 | PE-labeled | |||
| Lysozyme | LZ-2 | IgG1 | FITC-labeled | 50 tests | GIC-206 |
| Lactoferrin | 4C5 | IgG1 | PE-labeled | ||
| Myeloperoxidase | 8E6 | IgG1 | FITC-labeled | 50 tests | GIC-212 |
| Lactoferrin | 4C5 | IgG1 | PE-labeled | ||
| Myeloperoxidase | 8E6 | IgG1 | FITC-labeled | 50 tests | GIC-213 |
| CD3 | UCHT1 | IgG1 | PE-labeled | ||
| Myeloperoxidase | 8E6 | IgG1 | FITC-labeled | 50 tests | GIC-214 |
| CD22 | RFB4 | IgG1 | PE-labeled |
Caltag Medystems are the UK and Ireland distributor for Nordic Mubio products. If you have any queries, please do not hesitate to contact us, 01280 827460 or office@caltagmedsystems.co.uk.