V8 capillary


TITAN III Cellulose Acetate Electrophoresis


Alkaline Phosphatase Isoenzymes

Alkaline and Citrate Agar Hemoglobins



Acetate Membranes



Hemoglobin Controls

    Alkaline Phosphatase Indolyl Blue Reagent
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

    Alkaline Phosphatase Indolyl Blue Reagent
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

    Alkaline Phosphatase Indolyl Blue Reagent
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.



TITAN III and TITAN IV Hemoglobins

Cellulose Acetate & 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-Specific Stains

    Alkaline Phosphatase Indolyl Blue Reagent
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

    Alkaline Phosphatase Indolyl Blue Reagent
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).