Radioimmunoassay for tubulin detection

Guern, Christian Le; Pradelles, Philippe; Dray, F.

doi:10.1016/0014-5793(77)81065-x

Cited by 12 publications

(5 citation statements)

References 14 publications

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“…Binding of 1 2 5 1 -t~b~l i n to the antiserum was progressively inhibited by increasing amounts of unlabelled tubulin. Half-maximal inhibition (Iso) was obtained with 3.5 p g of tubulin, which is comparable to the radioimmunoassay of tubulin developed by Morgan et al (1978) and Bulinski et al (1980), but 70 times less sensitive than the assay described by Le Guern et al (1977), probably because of our mild iodination conditions. The sensitivity of the assay was approximately 0.4 pg, while the upper limit of the assay was approximately 25 pg.…”

Section: Radioimmunoassay Of Tubulinsupporting

confidence: 62%

Immunocytochemical Study of Microtubules in Chromaffin Cells in Culture and Evidence That Tubulin Is Not an Integral Protein of the Chromaffin Granule Membrane

Bader

Ciesielski-Treska

Thiersé

et al. 1981

Journal of Neurochemistry

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Bovine adrenal chromaffin cells were maintained in culture in Dulbecco's modified Eagle's medium containing 20% foetal calf serum and 10 units per ml of Nerve Growth Factor. Under these conditions, chromaffin cells developed up to five neurites per cell. The neurites showed lateral branches and varicosities along their trunk which ended with thick growth cone-like structures. Cultures of chromaffin cells were stained by indirect immunofluorescence with antibodies against (a) chromogranin A to follow the distribution of chromaffin granules, the catecholamine-storing organelles, and (b) tubulin, to study the microtubular system during outgrowth of neurites. Chromogranin A antibodies showed a very intensely staining punctate pattern, not randomly distributed but localized in neurites. Chromaffin granules were found to migrate from the cell body to reach neurite endings where they were densely packed. Intense staining was also observed in varicosities; a linear arrangement of granules was evident along neurite trunks. Tubulin antibodies decorated a complex network, clearly visible at the cell periphery and also in the growth cone-like structures, in the palm region of the growth cone. Colchicine treatment effected retraction of neurites and disappearance of organized microtubule networks; chromaffin granules were found in the perinuclear region of the cell. Some tubulin (0.2% of total membrane proteins) was found in the purified chromaffin granule membrane preparation; however, this tubulin is probably associated with contaminating plasma membranes. By the criteria of morphology and staining with antitubulin antibodies, adult bovine chromaffin cells in culture display characteristics similar to those of sympathetic neurones. In addition, they showed an exaggerated transport of granules. Adult bovine chromaffin cells in culture offer an excellent model for studying the role of microtubules and the contractile apparatus in relation to cell morphological changes and neurosecretion.

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Section: Radioimmunoassay Of Tubulinsupporting

confidence: 62%

Immunocytochemical Study of Microtubules in Chromaffin Cells in Culture and Evidence That Tubulin Is Not an Integral Protein of the Chromaffin Granule Membrane

Bader

Ciesielski-Treska

Thiersé

et al. 1981

Journal of Neurochemistry

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“…Covalent binding of the tubulin to bovine immunoglobulin, electrostatic coupling of tubulin to methylated bovine serum albumin, or mild dinitrophenylation did not lead to a significant enhancement of the antibody titers [60]. The antigen concentration can be lowered to 25pg per injection [82]. Another method of immunization involving tubulin cross-linked with glutaraldehyde to form an insoluble complex has been described [46].…”

Section: Preparation Of Antibodies Against Brain Tubulinmentioning

confidence: 99%

“…A more sensitive radioimmunoassay was developed by Le Guern ef al. [82], who iodinated the tubulin in the presence of chloramine-T and performed the radioimmunoassay in the presence of GTP. The radioimmunoassay was used to determine species-dependent immunological differences between vertebrate brain tubulin [97] as will be discussed subsequently.…”

Section: Quantitative Determination By Radioimmunoassaymentioning

confidence: 99%

Microtubule Protein: Tubulin

Gozes

Littauer

1982

Scand J Immunol

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“…Tubulin was purified from ham ster brain on a phosphocellulose column as described [8]. Purified actin was kindly provided by A.Olomucki (Collège de France, Paris).…”

Section: Methodsmentioning

confidence: 99%

“…The cells were fixed at 4,8,16, and 24 h and at 3, 4, 5 and 7 days after infection for immunocytological stainingand with 1.6% glutaraldehyde for conventional ultrastructural studies. Fixation for immunocytologi cal staining was slightly modified from the previous procedure [6]: the first 2-3 min of fixation with 6% paraformaldehyde at room temperature, and then 7-8 min of fixation in ice-cold water and 0.2% glutaral dehyde.…”

Section: Methodsmentioning

confidence: 99%

Localization with the Electron Microscope of p27 Rous Sarcoma Virus Internal Protein in Chicken Embryo Fibroblasts

Stanislawski¹

1983

Intervirology

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Protein 76 (Pr76), the precursor of the so-called ‘gag’ protein of avian leuko-sarcoma viruses, has been localized in the light and electron microscope by using the immunoperoxidase technique in Rous sarcoma virus-infected chick embryo fibroblasts. The kinetics of the progression of the protein through the cell was studied at different times after infection. The protein was first found (after 8 h) on ribosomes located near the nucleus and then proceeded rapidly to the periphery of the cell. After 5–6 days, intracytoplasmic staining faded away and only the ribosomes close to the plasma membrane were stained. A possible association of a protein with the cytoskeleton is suggested.

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Radioimmunoassay for tubulin detection

Cited by 12 publications

References 14 publications

Immunocytochemical Study of Microtubules in Chromaffin Cells in Culture and Evidence That Tubulin Is Not an Integral Protein of the Chromaffin Granule Membrane

Immunocytochemical Study of Microtubules in Chromaffin Cells in Culture and Evidence That Tubulin Is Not an Integral Protein of the Chromaffin Granule Membrane

Microtubule Protein: Tubulin

Localization with the Electron Microscope of p27 Rous Sarcoma Virus Internal Protein in Chicken Embryo Fibroblasts

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