PMID- 8491814
OWN - NLM
STAT- MEDLINE
DCOM- 19930616
LR  - 20190630
VI  - 613
IP  - 2
DP  - 1993 Apr 2
TI  - Enhanced sensitivity of immunoblotting with peroxidase-conjugated antibodies 
      using an adsorbed substrate method.
PG  - 281-8
AB  - Immunoblots probed with immunoglobulin E (IgE)-containing sera from allergic 
      patients are frequently used in allergy research. Current techniques for 
      detection of specific IgE include radiolabeled and enzyme-linked methods. 
      Although radiolabeled methods are very sensitive, many research groups prefer 
      non-radioactive procedures with equal or greater sensitivity. Alkaline 
      phosphatase (AP) and horse radish peroxidase (HRP) are the most frequently used 
      conjugating enzymes for immunoblotting with the former generally recognized as 
      more sensitive. We describe a method of immunoblot detection using HRP-conjugated 
      immunochemicals with sensitivity equal to and for some systems greater than that 
      of AP conjugates. An adsorbed substrate method for developing immunoblots probed 
      with HRP immunochemical conjugates is compared with traditional AP and HRP 
      methods. The adsorbed substrate system, when used to detect IgE binding to 
      allergic proteins, gives high resolution and delineates bands not otherwise seen. 
      The system has advantages of high sensitivity, rapid development and conservation 
      of immunochemicals. Problems of fading, sensitivity to heat and light, and high 
      background can be solved with increased washing, prompt photography and computer 
      scanning.
FAU - Barnes, C S
AU  - Barnes CS
AD  - Section of Allergy/Immunology, Children's Mercy Hospital, Kansas City, MO 64108.
FAU - Upadrashta, B
AU  - Upadrashta B
FAU - Pacheco, F
AU  - Pacheco F
FAU - Portnoy, J
AU  - Portnoy J
LA  - eng
GR  - R29 AI27476-01A1/AI/NIAID NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - Netherlands
TA  - J Chromatogr
JT  - Journal of chromatography
JID - 0427043
RN  - 0 (Antibodies)
RN  - 0 (Indicators and Reagents)
RN  - 37341-29-0 (Immunoglobulin E)
RN  - EC 1.11.1.- (Horseradish Peroxidase)
RN  - EC 3.1.3.1 (Alkaline Phosphatase)
SB  - IM
MH  - Adsorption
MH  - Alkaline Phosphatase/analysis/immunology
MH  - Alternaria/chemistry/growth & development
MH  - Antibodies/*immunology
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Horseradish Peroxidase/analysis/immunology
MH  - Humans
MH  - Immunoblotting/*methods
MH  - *Immunoenzyme Techniques
MH  - Immunoglobulin E/analysis/immunology
MH  - Indicators and Reagents
EDAT- 1993/04/02 00:00
MHDA- 1993/04/02 00:01
CRDT- 1993/04/02 00:00
PHST- 1993/04/02 00:00 [pubmed]
PHST- 1993/04/02 00:01 [medline]
PHST- 1993/04/02 00:00 [entrez]
AID - 10.1016/0378-4347(93)80143-r [doi]
PST - ppublish
SO  - J Chromatogr. 1993 Apr 2;613(2):281-8. doi: 10.1016/0378-4347(93)80143-r.