TITAN GEL Manual Agarose Electrophoresis

Alkaline Phosphatase Isoenzymes

 

TITAN GEL Alkaline Phosphatase Isoenzymes

The alkaline phosphatase (ALP) isoenzymes found in human serum originate from several sources with the greatest activity occurring in the bone, liver, intestine, and placenta. Because of wide distribution of alkaline phosphatase in tissue, limited information can be obtained from a total ALP assay. Fortunately, the tissue sources of elevated ALP in serum can be determined by identifying the isoenzyme.

The isoenzymes of alkaline phosphatase are unique in that some organs have only one major isoenzyme rather than multiple isoenzyme forms. The isoenzymes of ALP differ in their physicochemical and electrophoretic properties, and it is by taking advantage of these differences that individual isoenzymes can be identified. In addition to the liver, bone, intestinal and placental isoenzymes, macrohepatic, Regan, PA, Nagao, and renal isoenzymes have also been identified in serum.

The resolution, accuracy and convenience of the TITAN GEL Alkaline Phosphatase High Resolution procedure make it better than heat denaturation, isoelectric focusing and wheat germ lectin techniques for separating ALP isoenzymes.

TITAN GEL Alkaline Phosphatase Kit

Cat. No.	Item	Description
3058	TITAN GEL Hi Res Alk Phos	10 x 8 samples/gel
	Isoenzyme Kit

Kits include: 10 Gels, ALP Buffer, ALP Separation Enhancer, ALP Reagent, ALP Diluent, Blotter, Wicks, Templates.

The TITAN GEL high resolution agarose electrophoresis method for separation of alkaline phosphatase isoenzymes provides excellent resolution. Neuraminidase is used to enhance separation of bone and liver fractions. A unique additive to the gel matrix ensures separation of the macrohepatic fraction from bone and liver. The excellent sensitivity of the system allows detection and separation of ALP isoenzymes even in samples with low activity. Up to eight samples may be applied per gel.

Alkaline Phosphatase Isoenzyme Control

Cat. No.	Item	Description
5104	Gel ALP Isoenzyme Control	1 x 2 mL

This control is for use as a qualitative and/or quantitative control to aid in the identification of alkaline phosphatase isoenzymes by agarose gel electrophoresis. It is prepared from pooled human serum and contains liver and bone isoenzyme bands. The control is a stabilized liquid.

Also for use with REP and SPIFE assays.

CK Isoenzymes

 

TITAN GEL CK Isoenzymes

Electrophoresis is the reference method for assessing CK isoenzymes. There's practically no cross-reactivity and no false positives due to atypical isoenzymes or carryover from high total activity samples. Electrophoresis provides comprehensive information on CK-MM, CK-MB and CK-BB, resulting in greater accuracy in interpretation and efficiency in test performance. Resolution is excellent and test results, including scanning, are complete in less than one hour.

 

 

CK/LD Isoenzyme Control

Cat. No.	Item	Description
5134	CK/LD Isoenzyme Control	5 x 2 mL

Prepared with murine tissue extract and human serum, control is for quantitative and/or qualitative use in conventional agarose electrophoresis of CK and LD isoenzymes (3 CK and 5 LD isoenzymes present). Reconstituted control is stable 5 days at -20°C.
Also for use with SPIFE CK and LD and assays.
Remember: TITAN GEL Isoenzyme Incubation Chamber (Cat. No. 4062).

High Resolution Proteins

 

TITAN GEL High Resolution Proteins

The major advantage of high resolution electrophoresis over conventional protein electrophoresis is that it gives more information about the patient's condition. Fifteen or so proteins can be visualized by use of high resolution, high sensitivity agarose gel electrophoresis of serum. It is the method of choice for detecting monoclonal immunoglobulins in serum and urine. The superior resolution and sensitivity of the procedure separates and detects proteins with similar electrophoretic mobilities and those present in low concentrations that may be invisible on other methods.

 

 

TITAN GEL High Resolution Protein Kit

Cat. No.	Item	Description
3040	TITAN GEL Hi Res Protein Kit	10 x 7 samples/gel

Kits include: 10 Gels, HR Buffer, HR Stain, Blotters, Wicks, Templates.

This method is excellent for detecting monoclonal immunoglobulins in serum and urine and oligoclonal bands in CSF. The procedure requires less than one hour for results and eliminates the need for expensive cumbersome cooling systems. This kit is used when sufficient CSF sample is available for concentration to 80 fold. Seven samples may be run on each gel.

 

High Resolution Protein Marker

Cat. No.	Item	Description
5141	High Resolution Protein Marker	1 x 0.1 mL

A qualitative marker for use with the TITAN GEL, REP and SPIFE High Resolution Protein Systems. The marker exhibits multiple bands and simulates oligoclonal banding in CSF and serum. It provides assurance that sufficient resolution has been achieved and is a visual reference for differentiating oligoclonal bands. The marker can be stained with Hi Res Protein Stain or Silver Stain. The marker is a stabilized liquid prepared from human serum.

Also for use with REP and SPIFE Hi Res assays.

Remember: TITAN GEL Chamber Cooling Device (Cat. No. 3039), Amido Black Protein stain (an alternate to Coomassie Brilliant Blue Stain (Cat. No. 3048).

 

Immunofixation

 

TITAN GEL Immunofixation

Immunofixation electrophoresis (IFE) is characterized by its enhanced sensitivity, ease of interpretation, quick test results, and excellent resolution. IFE is a two stage procedure using agarose protein electrophoresis first, then immunoprecipitation. The specimen may be serum, urine or CSF. There are many applications for IFE but the primary use is characterization of paraproteins. Distinct protein separation, antigen/antibody specificity, and direct comparison of IFE bands with the corresponding protein pattern from the same system make interpretation easy and provide definitive results. Pre-cut wells on each antisera lane allow use of Titan Gel ImmunoFix Controls to provide verification of antisera reactivity on each patient gel.

 

TITAN GEL ImmunoFix 6-Channel Gels and Antisera

Cat. No.	Item	Description
3046	TITAN GEL ImmunoFix Kit	10 x 6 channels/gel
3046A	TITAN GEL ImmunoFix Gels	30 gels, no antisera

Kits include: 10 Gels, Acid Blue Stain, IFE Buffer, Sample Templates, Antisera Templates, Protein Fixative, Blotters, and 1 x 0.75 mL vial each of Antisera to IgG, IgA, IgM, Kappa and Lambda.

Each TITAN GEL ImmunoFix Kit contains reagents and supplies for performing 10 patient profiles on a standard 6-channel gel. The procedure requires just over an hour to complete. Wash and press steps require only 10 minutes. Antisera are color coded and packaged in applicator vials for easier application and quality assurance. Each gel contains wells for application of TITAN GEL IFE Controls.

TITAN GEL ImmunoFix-Plus 12-Channel Gels

Cat. No.	Item	Description
3067	TITAN GEL ImmunoFix-Plus	10 x 12 channels/gel

Kits include: 10 Gels, Acid Blue Stain, IFE Buffer, Blotters, Sample Templates, Antisera Templates.

The advantages of IFE are now enhanced by the versatility of TITAN GEL ImmunoFix-Plus. These gels have 12 channels instead of the traditional 6. This allows testing of two different patients while handling only one gel, or testing a patient's serum and urine on the same gel, or many other combinations. With this expanded gel and Helena's complete line of high affinity, high titer antisera, each laboratory can adapt IFE to meet its daily demands.

Each channel of the IFE-Plus Gel has application wells for TITAN GEL IFE Controls. Antisera and TITAN GEL IFE Control Kits are purchased separately.

TITAN GEL IFE Antisera Kit

Cat. No.	Item
3068	TITAN GEL IFE Antisera Kit

Kits include: Protein Fixative and 1 x 0.75 mL vial each of Antisera to IgG, IgA, IgM, Kappa and Lambda.

This kit provides the routine monospecific antisera needed for TITAN GEL ImmunoFix Procedures. The high affinity and titer are excellent for detecting and identifying paraproteins. The Protein Fixative provides a complete protein pattern for comprehensive work-up. Color-coded antisera are packaged in applicator vials.

IFE Antisera Kit II

Cat. No.	Item
3151	IFE Antisera Kit II

Kits include: Protein Fixative and 1 x 3 mL vial each of Antisera to IgG, IgA, IgM, Kappa Light Chains and Lambda Light Chains.
The Antisera Kit II provides adequate antisera for routine testing on 30 patient samples. The antisera are identical to those in Cat. No. 3068 but for the convenience of high volume users, vials have a greater fill volume. Labs ordering this antisera kit should also order an appropriate number of ImmunoFix gels only. Antisera are color coded and packaged in convenient applicator vials.

TITAN GEL IFE Kappa and Lambda Light Chain Antisera Kit

Cat. No.	Item
3069	IFE Kappa and Lambda Light Chain Antisera

Kits include: Protein Fixative and 2 vials each of Antisera to Kappa and Lambda.

The antisera kit is ideal for laboratories with a greater demand for kappa and lambda light chain antisera. These antisera react with both bound and free light chains. It is the same antisera contained in Cat. No. 3046 and Cat. No. 3068. For Free Light Chain Antisera (reacts with only free light chains) see Cat. No. 9412 and 9413. Packaged in convenient applicator vials.

TITAN GEL IFE Free Kappa and Free Lambda Antisera

Cat. No.	Item
9412	TITAN GEL IFE Free Kappa Antiserum, 1 x 1 mL
9413	TITAN GEL IFE Free Lambda Antiserum, 1 x 1 mL

TITAN GEL IFE Free Kappa and Free Lambda Antisera react only with the free light chains (not attached to the heavy chain portion of the immunoglobulin molecule) and can be used with both serum and urine. Packaged in convenient applicator vials.

TITAN GEL IFE Free Kappa and Free Lambda Antisera

Cat. No.	Item
9409	TITAN GEL IFE Antiserum to IgD, 1 x 1 mL
9410	TITAN GEL IFE Antiserum to IgE, 1 x 1 mL

When performing IFE, if a reaction occurs with kappa or lambda but no reaction occurs with the IgG, IgA, or IgM heavy chains, the patient sample should be retested with IgD and IgE antisera. A reaction indicates an IgD or IgE monoclonal gammopathy. Samples with a light chain reaction and no reaction with IgD or IgE are appropriately reported as Free Light Chain Disease. Packaged in convenient applicator vials.

TITAN GEL IFE Trivalent (IgG, IgA, IgM) Antiserum

Cat. No.	Item
9411	Trivalent Antiserum, 1 x 1 mL

TITAN GEL IFE Trivalent Antiserum contains antibodies to IgG, IgA, and IgM. It is useful as a screening antiserum for serum or urine, especially when testing serum and urine on the same patient. Monospecific IgG, IgA, and IgM can be used on the serum and Trivalent used on the urine. A heavy chain reaction in the urine can be assumed to be the same as the heavy chain reaction in the serum. When used with the 12-channel TITAN GEL IFE-Plus Gel, the protocol provides efficiency and economy in performing paraprotein studies.

TITAN GEL ImmunoFix Quality Control

Cat. No.	Item
9400	ImmunoFix Control Kit
	1 x 0.5 mL IgG Kappa Control
	1 x 0.5 mL IgA Lambda Control
	1 x 0.5 mL IgM Control

TITAN GEL ImmunoFix controls are used to confirm antisera reactivity on each patient gel. After electrophoresis, the appropriate control is applied to a well at the end of each channel on the gel. When antisera applied to the channels comes in contact with the control serum, an antigen/antibody reaction occurs, causing a precipitin ring to form around each well on the finished gel. To verify all phases of the procedure, the control can be treated as a patient sample and taken through every step of the procedure. The laboratory may elect to perform this quality control procedure on one gel from each lot of kits received.

Remember: TITAN GEL Isoenzyme Incubation Chamber (Cat. No. 4062), Dialamatic 1-10 μL Microdispenser (Cat. No. 6210), 1000 Staining Set (Cat. No. 5122, use with 3046)

 

 

 

Immunoelectrophoresis

 

TITAN GEL & Titan IV IEP

IEP is a reliable and accurate method for routine serum and urine protein evaluations, detecting both structural abnormalities and concentration changes. It has been widely used in identification of monoclonal gammopathies. Immunoelectrophoresis (IEP) combines two techniques, electrophoresis and immunodiffusion. In this two-part procedure, proteins are first separated by electrophoresis. Then antisera, complementary to the proteins under study, are applied to the gel and allowed to diffuse. When a favorable antigen to antibody ratio exists, a precipitin arc forms on the gel.

Both thick and thin gels are available for performing immunoelectrophoresis. The thick agarose, on a rigid plastic backing for ease of handling, requires a larger volume of antisera and patient samples, and forms a dense precipitin arc easily interpreted and photographed without staining the gel. The thin agarose gel, on a flexible plastic backing, provides economical use of antisera and lends itself to staining and drying of the gel for a permanent record.

 

TITAN GEL Immunoelectrophoresis

Cat. No.	Item
3047	TITAN GEL IEPlate Kit, 10 IEPlates plus blotters

TITAN GEL IEPlates provide fast, economical immunoelectrophoresis. The thin gel uses a reduced sample volume of 2 mL and antisera volume of 25 mL which means tremendous cost savings. Each gel has eight wells and seven troughs to accommodate a wide variety of antisera profiles. After an 18-hour incubation, non-precipitated proteins are washed from the gel. The gel can then be stained, destained and dried within one hour. The printed plastic backing makes sample and control identification easy and includes a preprinted scale for measuring migration distance.

Cat. No.	Item
9231	Antisera to Human IgA, Heavy Chain Specific, 1 x 2 mL
9232	Antisera to Human IgG, Heavy Chain Specific, 1 x 2 mL
9233	Antisera to Human Whole Serum, 1 x 2 mL
9234	Antisera to Human IgM, Heavy Chain Specific, 1 x 2 mL
9236	Antisera to Human (IgG, A, M) Trivalent, Heavy Chain Specific, 1 x 2 mL
9249	Antisera to Human IgD, Heavy Chain Specific, 1 x 2 mL
9250	Antisera to Human IgE, Heavy Chain Specific, 1 x 2 mL
9251	Antisera to Human (IgG, A, M, D, E) Pentavalent, Heavy Chain Specific, 1 x 2 mL
9257	Antisera to Human Lambda, LIght Chain Specific, 1 x 2 mL
9262	Antisera to Human Kappa, Light Chain Specific, 1 x 2 mL

Helena's antisera are produced in goats or sheep.

TITAN GEL IEP Stain

Cat. No.	Item
3049	TITAN GEL IEP Stain, 1 x 10 g

The stain is a Coomassie Brilliant Blue Stain in powder form designed especially for use with the TITAN GEL IEPlate Kit (Cat. No. 3047). Stain is dissolved in 1 L mixture of water, ethanol, and glacial acetic acid.

Remember:  IEP Sponge Wicks - Long (Cat. No. 9015), Dialamatic 1-10 μL Microdispenser (Cat. No. 6210), Electra B₁ Buffer (cat. No. 5016),  Normal Human Serum Control (Cat. No. 9010),  Albumin Marker (Cat. No. 9011).

TITAN IV IEPlates

Cat. No.	Item
9050	Titan IV IEPlates - 8 wells, 7 troughs, 5 gels/pkg
9061	Titan IV IEPlates - 7 wells, 6 troughs, 5 gels/pkg

Titan IV IEPlates are a thick agarose gel on a rigid plastic support. Each gel has a plastic lid and a reusable plastic pouch. Results may be interpreted without staining.

Cat. No. 9050 (see picture) has 8 wells and 7 troughs. Gels require 4 mL of sample and 75 mL of antisera. The well position is 17 mm from the cathode end of the trough. The recommended migration distance is 40 mm.

Cat. No. 9061 has 7 wells and 6 troughs. Gels require 4 mL of sample and 150 mL of antisera. The well position is 23 mm from the cathode end of the trough. The recommended migration distance is 20 mm. Precipitin arcs on this gel are slightly shorter and broader than those obtained with Cat. No. 9050.

 

Double Diffusion Plates

Cat. No.	Item
9009	Double Diffusion Plates, 10 gels/pkg

Gels contain 15 wells in three, 5-well configurations. Useful in determining antisera titer, antigen/antibody ratio, checking purity of antigen preparations, etc.

 

Gel Trays

Cat. No.	Item
5009	Gel Trays, 76 x 102 mm, 20/pkg
5010	Gel Trays, 25 x 76 mm, 20/pkg

The 76 x 102 mm (3" x 4") gel tray is designed to be filled with 12.5 mL of liquid agarose. The 25 x 76 mm (1" x 3") trays require a 2.9 mL volume.

IEP Normal Human Serum Control and Albumin Marker

Cat. No.	Item
9010	Normal Human Serum Control, 1 x 2 mL
9011	Albumin Marker, 1 x 2 mL

The control is a stabilized liquid prepared from pooled normal human serum for use as a normal control serum for immunoelectrophoresis procedures. Albumin Marker is bromphenol blue which binds to albumin. The marker is added to the Normal Human Serum Control so the control can be used as a migration marker in immunoelectrophoresis procedures.

Remember:  IEP Sponge Wicks - Long (Cat. No. 9015),  Electra B1 Buffer (Cat. No. 5016).

LD Isoenzymes

 

 

TITAN GEL LD Isoenzymes

Different tissues contain varying concentrations of the individual LD isoenzymes: heart muscle contains LD1, liver and skeletal muscle contain LD5.

Electrophoresis, the method of choice for LD isoenzyme analysis, provides quantitation of all five isoenzymes in one test procedure. Electrophoresis accurately quantitates the LD1 and LD2 isoenzymes, as well as provides meaningful information on potential complications and secondary disease states, particularly pulmonary infarction and hepatic congestion.

 

 

 

CK/LD Isoenzyme Control

Cat. No.	Item	Description
5134	CK/LD Vis Isoenzyme Control	5 x 2 mL

Prepared with murine tissue extract and human serum, control is for quantitative and/or qualitative use in conventional agarose electrophoresis of CK and LD isoenzymes (3 CK and 5 LD isoenzymes present). Reconstituted control is stable 5 days at -20°C.

Also for use with SPIFE CK and LD assays.

Remember: TITAN GEL Isoenzyme Incubation Chamber (Cat. No. 4062).