TITAN IV Agar Electrophoresis
Titan IV 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).
Titan IV Citrate Hemoglobins
Titan® III and Titan IV® Hemoglobins
Cellulose Acetate and Citrate Agar
Numerous hemoglobin variants, many capable of producing severe hemolytic anemia and even death, have been identified. This has led to widespread implementation of neonatal screening programs. Counseling of individuals or marital partners, regarding the risk of having children with abnormal hemoglobins, is also common. Early detection of certain abnormal hemoglobins can alert parents to possible problems before onset of symptoms and allow physicians to better manage patients with hemoglobin variants.
Electrophoresis is generally considered the best method for separating and identifying abnormal hemoglobins. Via a two-step process, blood samples are initially screened using
alkaline hemoglobin electrophoresis, followed by confirmation of abnormals
by electrophoresis at an acid pH.
Helena hemoglobin procedures are widely used for hemoglobin screening programs. Many samples can be handled quickly, easily and cost-effectively. Interpretation is made by comparing the patient's hemoglobin pattern to a control sample.
Cellulose Acetate Assay Time: approximately 40 minutes.
Citrate Agar Assay Time: approximately 55 minutes.
See also:
SPIFE Alkaline and Acid Hemoglobin Electrophoresis methods.
QuickGel Alkaline and Acid Hemoglobin Electrophoresis .
Hemoglobin Controls
| Cat. No. | Item |
| 5328 | AA2 Hemo Control, 1 x 1 mL |
| 5329 | ASA2 Hemo Control, 1 x 1 mL |
| 5330 | AFSA2 Hemo Control, 1 x 1 mL |
| 5331 | AFSC Hemo Control, 1 x 1 mL |
Hemo Controls are stabilized liquids prepared from fresh-packed human red blood cells hemolyzed with Hemolysate Reagent. They are for qualitative and quantitative use in identification and quantitation of hemoglobins. AFSC Hemo Control is recommended for use with citrate agar.
Hemolysate Reagents
| Cat. No. | Item |
| 5125 | Hemolysate Reagent, 1 x 250 mL |
| 5127 | Hemolysate Reagent w/o KCN, 1 x 250 mL |
| 5128 | Hemolysate Reagent (Powder), 1 x 1.8 g |
This reagent hemolyzes red blood cells and stabilizes hemoglobin in one easy step. It eliminates many tedious, time-consuming steps in preparing hemolysates. The reagent contains 0.005 M EDTA with 0.07% potassium cyanide to assist in stabilizing the hemoglobin. The powdered reagent is dissolved in 1.0 liter deionized water.
Titan IV Citrate Agar and Citrate Buffer for Hemoglobins
| Cat. No. | Item |
| 2400 | Titan IV Citrate Agar, 5/pkg |
| 5121 | Citrate Buffer, 10 x 15.5 g |
The lower pH of the Titan IV Citrate Agar allows it to be used as a primary screening procedure for detection of HbS in newborns and as a confirmation of abnormal hemoglobins detected by alkaline electrophoresis. Up to 14 samples plus two controls can be run on each plate. Samples are easily applied with Zip Zone Applicator (Cat. No. 4080). Citrate Buffer is used as the chamber buffer for the citrate agar procedure.
Hemoglobin-Specific Stains
| Cat. No. | Item |
| 5036 | o-Dianisidine, 1 x 2 g |
| 5041 | o-Tolidine, 1 x 2 g |
o-Dianisidine is a sensitive, hemoglobin-specific stain used for staining hemoglobins separated on Titan IV Citrate Agar. An alternate stain for use with Titan IV Citrate Agar, o-Tolidine yields blue-colored hemoglobin bands. Both stains are powders that are dissolved in 1.0 liter methanol for use.
Titan III-H Cellulose Acetate and Supre-Heme Buffer for Hemoglobins
| Cat. No. | Item |
| 3021 | Titan III-H Cellulose Acetate, 94 x 76 mm |
| 3022 | Titan III-H Cellulose Acetate, 60- x 76 mm |
| 5802 | Supre-Heme Buffer, 10 x 14.6 g |
Titan III-H Cellulose Acetate membranes are for use in the alkaline hemoglobin electrophoresis. Membranes are available in two sizes and packaged 25 per box. Supre-Heme Buffer is a Tris-EDTA-Boric Acid buffer used in the alkaline hemoglobin procedure.
Complete the Cellulose Acetate and Citrate Agar Hemoglobin Procedures with these supplies: Ponceau S Stain (Cat. No. 5526), Clear Aid (Cat. No. 5005), and Titan IV Aligning Base (Cat. No. 4083).
Hemoglobin F Immunodiffusion Kit
| Cat. No. | Item |
| 9325 | HbF QUIPlate Kit, 3 x 16 wells |
The HbF QUIPlate Kit is for quantitation of Hemoglobin F by radial immunodiffusion. RID is much simpler and requires much less hands-on time than cumbersome alkali denaturation methods, which are also unreliable at low levels of HbF. The reference standards provided with the kit ensure accurate and precise quantitation from 0.5% to 10% HbF and are prepared from lyophilized hemoglobin. Samples with >10% HbF should be diluted with HbF Diluent. The crisp, well-defined precipitin rings can be read at 24 hours. Kit contains three 16-well plates, plus standards and graph paper for preparing a standard curve.
HbF QUIPlate Control and Diluent
| Cat. No. | Item |
| 9340 | HbF QUIPlate Control, 1 x 0.2 mL |
| 9341 | HbF QUIPlate Diluent, 1 x 2.0 mL |
HbF Control is a stabilized liquid prepared from human cord blood. It is used to verify plate/curve stability, allowing re-use of the standard curve for up to 28 days. HbF Diluent is prepared from purified human hemoglobin, providing a diluent completely free of HbF for diluting patient samples with >10% HbF.
Complete the QUIPlate procedure with: Dialamatic 1-10 mL Microdispenser (Cat. No. 6210).