Chemiluminescent image detection of haptoglobin phenotyping after polyacrylamide gel electrophoresis.
The development of an enhanced chemiluminescence detection method for the rapid detection of haptoglobin phenotyping after polyacrylamide gel electrophoresis is described in this paper. The enhanced chemiluminescence detection is based upon chemiluminescent reaction between luminol and hydrogen peroxide. Increased sensitivity and dynamic range are achieved by employing ammonium persulfate to enhance the chemiluminescence signal. Detection of haptoglobin phenotypes in human blood serum was easily achieved even without the addition of hemoglobin. Different polyacrylamide gel electrophoresis results were found between pure serum and hemoglobin-supplemented serum. Applying the suggested enhanced chemiluminescence detection, the original combining forms of haptoglobin and hemoglobin can be detected. The linear range of haptoglobin is 0.1-13.3 microg/mL, with a detection limit of 1.21 ng (sample loading volume 15 microL). Other proteins, such as catalase and ferritin, can also be detected using enhanced chemiluminescence detection. All detections after polyacrylamide gel electrophoresis were completed within 15 min. The proposed detection is very fast, compared to traditional methods using staining detection (minutes versus hours).
Huang
Guangming
G
Department of Chemistry, Beijing Normal University, Beijing 100875, PR China.
Ouyang
Jin
J
Delanghe
Joris R
JR
Baeyens
Willy R G
WR
Dai
Zhongxin
Z
D000645:Ammonium Sulfate / Q000737:chemistry / Q000494:pharmacology; D002482:Cellulose / Q000031:analogs & derivatives* / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006242:Haptoglobins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D006454:Hemoglobins / Q000737:chemistry / Q000378:metabolism; D008163:Luminescent Measurements / Q000379:methods*; D010641:Phenotype; D013997:Time Factors
D006242:Haptoglobins; D006454:Hemoglobins; C031276:ammonium peroxydisulfate; C005062:acetylcellulose; D002482:Cellulose; D000645:Ammonium Sulfate