|
|
|
|
 |
|||
|
|||
Additional factors influencing sensitivity in the tetramethyl benzidine method for horseradish peroxidase neurohistochemistry
MM Mesulam, E Hegarty, H Barbas, KA Carson, EC Gower, AG Knapp, MB Moss and EJ Mufson
In experiments that use horseradish peroxidase (HRP) and tetramethyl benzidine (TMB) for tracing neural connections, the activity of tissue- bound enzyme as well as the stability of the resultant reaction product are influenced by the duration of storage, the composition of the storage medium, the type of counterstaining and even the details of histological dehydration. Furthermore, the conditions for preserving HRP activity are very different from those necessary for preserving the stability of the tetramethyl benzidine (TMB) reaction product. Thus, tissue-bound HRP activity is stable at a neutral pH, while a much lower pH, around 3.3, is required for preserving the stability of the TMB reaction product. Recent evidence indicates that the stabilization bath in sodium nitroferricyanide that was previously recommended is not necessary. However, gradual dehydration of mounted sections is essential for long-term stability. Excessive counterstaining and excessive dehydration interfere with the detection of reaction product. These considerations are pertinent to experiments using free HRP as well as to those where the enzyme has been conjugated to wheat germ agglutinin.
Volume 28,
Issue 11,
pp. 1255-1259,
11/01/1980
Copyright © 1980 by The Histochemical Society
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati 
Twitter What's this?
This article has been cited by other articles:
|
|
 |
|
  N. Salichon, P. Gaspar, A. L. Upton, S. Picaud, N. Hanoun, M. Hamon, E. De Maeyer, D. L. Murphy, R. Mossner, K. P. Lesch, et al. Excessive Activation of Serotonin (5-HT) 1B Receptors Disrupts the Formation of Sensory Maps in Monoamine Oxidase A and 5-HT Transporter Knock-Out Mice J. Neurosci., February 1, 2001; 21(3): 884 - 896. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Komatsu, M. Kinoshita, and I. Murakami Neural Responses in the Retinotopic Representation of the Blind Spot in the Macaque V1 to Stimuli for Perceptual Filling-In J. Neurosci., December 15, 2000; 20(24): 9310 - 9319. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. I. Romero, M. A. Romero, and G. M. Smith Visualization of Axonally Transported Horseradish Peroxidase Using Enhanced Immunocytochemical Detection: A Direct Comparison with the Tetramethylbenzidine Method J. Histochem. Cytochem., February 1, 1999; 47(2): 265 - 272. [Abstract] [Full Text]
|
|
|
|
 |
|
 R. Kalb Regulation of motor neuron dendrite growth by NMDA receptor activation Development, January 11, 1994; 120(11): 3063 - 3071. [Abstract] [PDF]
|
|
| Guidelines | Subscriptions | About | exPRESS - Current - Archive | Business Information | Contact |
