IgG IEF

The Helena Biosciences SAS IgG IEF kit is intended for the identification of IgG-specific oligoclonal banding in serum and cerebro-spinal fluid (CSF) by agarose gel isoelectric focusing and immunoblotting. This technique is considered to be the “Gold Standard”¹ method for the determination of intrathecal IgG synthesis in the clinical diagnosis of multiple sclerosis and offers advantages over other methods such as IgG quotient, blood-CSF barrier function, direct staining and in-gel fixation methods. Many studies have shown that direct silver staining techniques exhibit reduced sensitivity and specificity to IgG.

Gold Standard method

Isoelectric Focusing and IgG immunodetection, the most sensitive paraclinical method for use in conjunction with MRI²
Up to 10 paired serum CSF samples per gel
No pre-concentration of samples required
Simple procedure, CE marked
The most sensitive procedure for humoral immune responses / oligoclonal band in multiple sclerosis detection¹
Optimised protocol and ampholytes for fully reproducible results — allowing the avoidance of “home brew” failures
Specific for IgG, no interference from other protein bands
Clear homogenous background, allows clear identification of negative samples
Kappa and Lambda antibodies available for easier interpretation of difficult samples
IgG IEF Control available
Can be used on multiple SAS platforms
All required components provided in ready-to-use format

Having dealt with Helena for 20 years now, I am obviously convinced of their excellence. The analysis of spinal fluid has been a Cinderella area due to a combination of smaller numbers of samples yet higher levels of sophistication required to detect oligoclonal IgG patterns. Helena have maintained the highest levels of quality control. This culminated in the recognition by the US Food and Drugs Administration of their kit for spinal fluid analysis. Thus the technique was further adopted by an International group of experts on the laboratory diagnosis of Multiple Sclerosis. It remains the “gold standard” in this field. Professor E J Thompson PhD MD DSc FRCPath FRCP,
The National Hospital for Neurology & Neurosurgery

Available gel sizes

Small

SAS IgG IEF Kit

REF:	102200
Tests per gel:	5
Gels per kit:	10
Compatibility:	SAS-MX IEF chamber, SAS-1+, SAS-3, SAS Vitrési

SAS-MX IEF

SAS IgG IEF Plus Kit

REF:	104200
Tests per gel:	10
Gels per kit:	10
Compatibility:	SAS-MX IEF chamber, SAS-3

Oligoclonal bands and Isoelectric Focusing

Oligoclocal banding analysis is used in combination with MRI scans to confirm Multiple Sclerosis when the situation is still clinically uncertain. The presence of oligoclonal bands in cerebro-spinal fluid (CSF) is indicative of intrathecal IgG production. This indicates an immune response to the central nervous system which is characteristic of Multiple Sclerosis.

The Helena Biosciences oligoclonal banding assay uses the Isoelectric Focusing technique (IEF). CSF-derived proteins are separated in the gel via a pH gradient which causes charged proteins to move up to the point at which their net electrical charge is neutral — known as the proteins’ Isoelectric Point (pI).

IEF in combination with immunoblotting allows the Helena Biosciences IgG method to provide clear, precise oligoclonal band detection. Our simple, step-by-step procedure provides accurate and efficient diagnosis.

Example membrane appearances

Immunoblotting

Immunoblotting is one of the most sensitive methods for the detection of proteins following electrophoresis.

Combined with IEF, immunoblotting is the “Gold Standard” method for detecting oligoclonal bands
The use of other methods may result in mis-diagnosis
The use of quantitative assays is not equivalent to qualitative immunoblotting²
Immunoblotting gives an unsurpassed dynamic range of three orders of magnitude

References

Freedman et al. “Recommended Standard of Cerebrospinal Fluid Analysis in the Diagnosis of Multiple Sclerosis.” Arch Neurol/Vol 62, June 2005. Andersson et al. “Cerebrospinal Fluid in the Diagnosis of Multiple Sclerosis: A Consensus Report.” Journal of Neurology, Neurosurgery and Psychiatry 1994; 57:897-902.
McDonald WI et al. “Recommended diagnosis criteria for multiple sclerosis: guidelines from the International Panel on the Diagnosis of Multiple Sclerosis”. AnnNeurol. 2001; 50: 121-127.
Verbeek et al. “Comparison of Methods for the Detection of Oligoclonal IgG Bands in Cerebrospinal Fluid and Serum: Results of the Dutch Quality Control Survey”. Clin Chem. 2002; 48: 1578-1580.

Catalogue numbers

In addition to the gel kits, Helena Biosciences offers a control designed to ensure peak performance of all stages of the process from focusing to protein transfer to immunodetection.

IgG IEF
102200	SAS IgG IEF Kit
104200	SAS IgG IEF Plus Kit
Optional Antisera
102206	SAS Kappa IEF Kit
102207	SAS Lambda IEF Kit
102208	SAS Free Kappa IEF Kit
102209	SAS Free Lambda IEF Kit
Control
102210	IgG IEF Control