|
|
|
|
 |
|||
|
|||
An indirect immunofluorescence procedure for staining the same cryosection with two mouse monoclonal primary antibodies
SA Lewis Carl, I Gillete-Ferguson and DG Ferguson
Department of Physiology and Biophysics, University of Cincinnati, College of Medicine, Ohio 45267-0576.
Indirect immunofluorescence procedures reported thus far are not effective at localizing two antigens in the same preparation when both primary antibodies are raised in the same species. In this case, the secondary antibodies can crossreact with both primary antibodies. We report here a protocol in which mouse monoclonal antibodies (MAb) specific for actin and myosin were used sequentially to stain the same frozen section of guinea pig skeletal muscle. The myosin-specific mAb was applied first and was localized with a rabbit anti-mouse IgG- rhodamine secondary antibody. The sections were then "blocked" with a non-binding mouse MAb and unconjugated goat anti-mouse IgG F(ab) fragments. The actin-specific mAb was then applied and localized with a rabbit anti-mouse IgG-fluorescein secondary antibody. Laser scanning confocal microscopy and image analysis demonstrated that the I-bands, the A-bands, and the H-bands of each sarcomere were clearly identifiable by this approach. This protocol is not limited to use with mouse MAb but can be easily modified to permit indirect immunolocalization of two antigens in the same sample using any pair of same-species primary antibodies.
Volume 41,
Issue 8,
pp. 1273-1278,
08/01/1993
Copyright © 1993 by The Histochemical Society
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  M. A. Gomez-Climent, E. Castillo-Gomez, E. Varea, R. Guirado, J. M. Blasco-Ibanez, C. Crespo, F. J. Martinez-Guijarro, and J. Nacher A Population of Prenatally Generated Cells in the Rat Paleocortex Maintains an Immature Neuronal Phenotype into Adulthood Cereb Cortex, October 1, 2008; 18(10): 2229 - 2240. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. M. Baleato, P. L. Guthrie, M.-C. Gubler, L. K. Ashman, and S. Roselli Deletion of Cd151 Results in a Strain-Dependent Glomerular Disease Due to Severe Alterations of the Glomerular Basement Membrane Am. J. Pathol., October 1, 2008; 173(4): 927 - 937. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. Merigo, D. Benati, M. Galie, C. Crescimanno, F. Osculati, and A. Sbarbati Immunohistochemical Localization of Cystic Fibrosis Transmembrane Regulator and Clara Cell Secretory Protein in Taste Receptor Cells of Rat Circumvallate Papillae Chem Senses, March 1, 2008; 33(3): 231 - 241. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. M. H. Ravindranath, A. Devarajan, and T. Uchida Spatiotemporal Expression of Ameloblastin Isoforms during Murine Tooth Development J. Biol. Chem., December 14, 2007; 282(50): 36370 - 36376. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. Borzacchiello and F. Roperto Double-labeling immunofluorescence: an alternative to detect multiple antigens in tissue sections. Vet. Pathol., January 1, 2006; 43(1): 83 - 83. [Full Text] [PDF]
|
|
|
|
 |
|
 H. Ino Application of Antigen Retrieval by Heating for Double-label Fluorescent Immunohistochemistry with Identical Species-derived Primary Antibodies J. Histochem. Cytochem., September 1, 2004; 52(9): 1209 - 1217. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. K. Brown, A. D. Pemberton, S. H. Wright, and H. R.P. Miller Primary Antibody-Fab Fragment Complexes: A Flexible Alternative to Traditional Direct and Indirect Immunolabeling Techniques J. Histochem. Cytochem., September 1, 2004; 52(9): 1219 - 1230. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Heilbronn, M. Engstler, S. Weger, A. Krahn, C. Schetter, and M. Boshart ssDNA-dependent colocalization of adeno-associated virus Rep and herpes simplex virus ICP8 in nuclear replication domains Nucleic Acids Res., November 1, 2003; 31(21): 6206 - 6213. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Uchihara, A. Nakamura, H. Nakayama, K. Arima, N. Ishizuka, H. Mori, and S. Mizushima Triple Immunofluorolabeling with Two Rabbit Polyclonal Antibodies and a Mouse Monoclonal Antibody Allowing Three-dimensional Analysis of Cotton Wool Plaques in Alzheimer Disease J. Histochem. Cytochem., September 1, 2003; 51(9): 1201 - 1206. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. A. Haxhiu, P. Kc, B. Neziri, B. K. Yamamoto, D. G. Ferguson, and V. J. Massari Catecholaminergic microcircuitry controlling the output of airway-related vagal preganglionic neurons J Appl Physiol, May 1, 2003; 94(5): 1999 - 2009. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. A. Haxhiu, B. K. Yamamoto, I. A. Dreshaj, and D. G. Ferguson Activation of the midbrain periaqueductal gray induces airway smooth muscle relaxation J Appl Physiol, August 1, 2002; 93(2): 440 - 449. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. Brouns, L. Van Nassauw, J. Van Genechten, M. Majewski, D. W. Scheuermann, J.-P. Timmermans, and D. Adriaensen Triple Immunofluorescence Staining with Antibodies Raised in the Same Species to Study the Complex Innervation Pattern of Intrapulmonary Chemoreceptors J. Histochem. Cytochem., April 1, 2002; 50(4): 575 - 582. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Matsushita, M. Oyamada, H. Kurata, S. Masuda, A. Takahashi, T. Emmoto, I. Shiraishi, Y. Wada, T. Oka, and T. Takamatsu Formation of Cell Junctions Between Grafted and Host Cardiomyocytes at the Border Zone of Rat Myocardial Infarction Circulation, November 9, 1999; 100(90002): II-262 - 268. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. M. McGowan and P. A. Coulombe Onset of Keratin 17 Expression Coincides with the Definition of Major Epithelial Lineages during Skin Development J. Cell Biol., October 19, 1998; 143(2): 469 - 486. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Q. L. Lu and T. A. Partridge A New Blocking Method for Application of Murine Monoclonal Antibody to Mouse Tissue Sections J. Histochem. Cytochem., August 1, 1998; 46(8): 977 - 984. [Abstract] [Full Text]
|
|
| Guidelines | Subscriptions | About | exPRESS - Current - Archive | Business Information | Contact |