Contact Us
Sign In / Sign Up for FREE
Go to advanced search...

Immunolocalization of the microtubule cytoskeleton - Essay Example

Comments (0) Cite this document
Proteins are probably the most diverse substances found in the cells found in both the single and multicellular organisms. Estimated around 10 000 different proteins, each protein has its specific function. Cellular functions are often localized in specific compartments of the cell, and being able to localize the unknown proteins may provide important information in determining the function of the proteins…
Download full paperFile format: .doc, available for editing
GRAB THE BEST PAPER92% of users find it useful
Immunolocalization of the microtubule cytoskeleton
Read TextPreview

Extract of sample "Immunolocalization of the microtubule cytoskeleton"

Download file to see previous pages The most prominent methods are: Western blot, spectrophotometry, enzyme assay, immunoprecipitation and immunostaining. In immunostaining, the method used during this procedure, an antibody is used to detect a specific protein epitope. These antibodies can be monoclonal or polyclonal. Then enzymes such as horseradish peroxidase or alkaline phosphatase are commonly used to catalyse reactions that give a coloured or chemiluminescent product. Fluorescent molecules can be visualised using fluoresence microscopy.
During this experiment, the distribution of the protein tubulin in normal rat kidney (NRK) cells is explored. A monoclonal antibody that is specific for the -subunit of tubulin is used. Tubulin polymerizes into long, 25-nm wide microtubules that we will visualize with tubulin antibodies. The formation and maintenance of microtubules is cold sensitive. At 4C, microtubules are destabilized and they depolymerize. At 37C, human body temperature, they remain polymerized. Photograph 1 shows cells incubated at 4 0C, while photograph 2 shows cells incubated at 37 0C. At this stage of the procedure, microtubules cannot be detected in either of the photographs.
3 separate plates are used to create the NRK cells culture. ...
The coverslip is then immersed in 3.7% formaldehyde (in PBS) at room temperature. Immersion in 0.2% Triton-X 100 (in PBS) detergent at room temperature follows. To prevent the subsequent antibody from sticking non-specifically to the cells on the coverslip, the coverslips should be incubated with cell-side-up with a 100-l droplet of 3% (w/v) bovine serum albumin (BSA). The incubation is done on a piece of Parafilm in a humid Petri dish for 15 minutes. An anti-tubulin dilution is prepared with 1 part antibody + 75 parts 1% (w/v) BSA in PBS. A 200 l total of antibody is necessary for the three coverslips. This tubulin antibody is a monoclonal antibody (12G10) and recognizes the -subunit of tubulin. Then each coverslip is overlayed each with a 100-l droplet of the diluted tubulin antibody. After that, a secondary antibody with 1 + 200 in 1% (w/v) BSA in PBS is used. The coverslips are placed back into the Petri dish chamber and overlay with a 100-l droplet of secondary antibody and they are incubate for 30 minutes at 37C. Therefore, we have 3 coverslips, all treated with the secondary antibody, 2 of them are treated with the primary antibody (one of them is incubated at 37C, and the other incubated at 4C) and 1 that is not treated with the primary antibody (incubated at 37C). The coverslips then are placed back into the Petri dish chamber and are overlaid for 30 minutes at 37C with a100-l droplet of Vectastain ABC reagent. This reagent contains a complex of avidin and the enzyme horseradish peroxidase (HRP). They are washed again with PBS. The drops of DAB (diaminobenzidine; a carcinogenic compound) are applied to the coverslips and incubate at room temperature for 5 minutes. After DAB incubation, ...Download file to see next pagesRead More
Cite this document
  • APA
  • MLA
(“Immunolocalization of the microtubule cytoskeleton Essay”, n.d.)
Immunolocalization of the microtubule cytoskeleton Essay. Retrieved from
(Immunolocalization of the Microtubule Cytoskeleton Essay)
Immunolocalization of the Microtubule Cytoskeleton Essay.
“Immunolocalization of the Microtubule Cytoskeleton Essay”, n.d.
  • Cited: 0 times
Comments (0)
Click to create a comment or rate a document

CHECK THESE SAMPLES OF Immunolocalization of the microtubule cytoskeleton


.... The family is made of transmembrane domains that have a general structure of the hydrophobic helix. It is surrounded by amphipathic helicies. Most of the action for this family is on the mitochondrial membrane (Wayne, 57) Apoptosis is also affected by cytochrome. Cytochromes are membrane bound homoproteins containing the heme groups and carry out electron transport. They are mainly within monomeric proteins e.g. cytochrome c family. They carry out oxidation and reduction in a sequence for efficiency. Cytochrome c is a protein that is very soluble at 100g/l. It is involved in the initiation of apoptosis (Geoffrey and Cooper, 77) Similarly important in apoptosis are the microtubules. They are part of the cytoskeleton. They grow to 25...
4 Pages(1000 words)Assignment

Microtubule Composition and Function in Eukaryotic Systems

...? MICROTUBULE COMPOSITION AND FUNCTION IN EUKARYOTIC SYSTEMS Word count: 1013 (references and figure captions excluded) Microtubule Composition and Function in Eukaryotic Systems Eukaryotic cells are differentiated from prokaryotic cells from the presence of membrane-bound organelles, an organized nuclear region, and a cytoskeleton network that preserves the shape of the cell. The cytoskeleton and its network in particular have integral roles in maintaining the cell’s shape, facilitating in cytoplasmic streaming, as well as in the movement and arrangement of the organelles within the cell especially during cell division (Galjart, 2010). The cytoskeleton is composed of three kinds of connective fibers, the smallest...
4 Pages(1000 words)Essay

Application of Microscopy in Biomedical Sciences

New applications are used to find the path of the unexpected discoveries (Suhling, French and Pillips 2004). According to Haupt, Pelling and Horton (2006) the detection of material subjected to damage the cells is a valuable strategy for both researchers and manufacturers of scientific equipments. A novel assay is required to study the Immunostaining and analysis of adherent cells by using the microscopic developments. This study reveals the analysis of adherent analysis by the treatments of inhibitors and other important fluorescence material (Vincent and Maiese 1999). Khosarvi-Fa et al (2008) determined the fixative solution used on the monolayer for the adherent cells DeMedeiore and Aho (2009) introduced the detection of inter...
6 Pages(1500 words)Lab Report

Molecular biology of the neuronal cytoskeleton

...? [Teacher’s Biology Molecular biology of the neuronal cytoskeleton. The cytoskeleton is a key defining component in eukaryotic cells. Most people refer to the cytoskeleton as a single component; however, it is comprised of three fundamental structures (Wayne and Morris, n.d). These include the microtubules, microfilaments and neurofilaments, they have different functions, but maintain interaction with one another (Brady and Kirkpatrick, 1). These fibrous subcomponents are synthesized from the polymerization of certain protein subunits. They each have different protein subunit compositions which are determined by the cell type or sub domain (Brady and Kirkpatrick, 1). These structures play a fundamental role in maintaining the form...
3 Pages(750 words)Article


... in the cardiac cells also function through microtubule assembly and its binding with different accessory proteins (Nicastro et al. 2006). Lastly, the topical review of Okamoto and Forte (2001) indicates the functional significance of such associations in vesicular trafficking machinery, present almost in all cells. This occurs through recruitment and recycling of specific membrane proteins assigned to perform specific functions. In the context of gastric parietal cells and the process of hydrochloric acid secretion, they contend that microtubules located in the apices of the parietal cells bind with H+K+-ATPase leading to remodeling of the apical membranes which begins a cascade of reactions between cytoskeleton and the vesicular...
6 Pages(1500 words)Essay


...) endocytosis of junctional proteins. Again, these mechanisms need further exploration but the existing understanding is that reorganization of F-actin causes destabilization of transinteraction between TJ and AJ proteins of the adjacent epithelial cells and in turn initiate AJC internalization. Microtubules, which are also an important element of intracellular cytoskeleton have also been found to be closely related to the cell movement as a whole or any intracellular structure formation like, vesiscles and their movement. Data support the abundance of microtubules in differentiated renal and intestinal cells and depolymerization of microtubules leading to disruption of the integrity of TJs and Ajs in some tissues like lungs...
6 Pages(1500 words)Essay

Red Blood Cells Cytoskeleton

... membrane. In the membrane cytoskeleton, spectrin is the most commonly found protein, forming long and elastic heterodimers, that join head to head to form heterotetramers, the tails of which are composed of F-actin, protein 4.1, and actin-binding proteins called dematin, adducin, tropomyosin, and tropomodulin (Stokes). This whole meshwork of proteins is attached to the cytoskeleton at two places: “one mediated by ankyrin that couples spectrin to Band 3 and the other mediated by protein 4.1 that couples the junctional complex to Glycophorin C” (Stokes, para.2). So, we see that human red blood cells are deficient in actin–myosin–microtubule cytoskeleton, which is helpful in cell shape determination, according to a recent research (Acton 69...
1 Pages(250 words)Article

Application of Microscopy in Biomedical Sciences

New applications are used to find the path of the unexpected discoveries (Suhling, French and Pillips 2004). Atomic Force Microscopy is the most important technique used in the biomedical applications, but it cannot compete with the traditional electron microscopy and confocal microscopy because of speeds at which later techniques capture the images (Haupt, Pelling and Horton, 2006).
Some other scholars also emphasized on the importance of confocal microscopy used for the observation of living cells. Use of the confocal microscopy is now common as non-ionizing radiations are employed, which are also used for the tissue preparation and study of the living cells. TGFβ stands for the transforming growth factor β, a family of the...
6 Pages(1500 words)Lab Report


This interaction, revealed by the structure, is branded the EB1-like motif and there is possibility that the binding interaction of CLIP family depends on this. Intuitively, considering that CLIP experiences binding at the acidic tail of tubulin, it is also possible that the acidic tail of EB1 also provides the same condition allowing for binding. 12
One of the important parts of the cell micro skeleton is the microtubules. The dynamics of microtubules is a very significant feature of cell control and cell morphogenesis. According to research carried out recently, microtubules are attributed with the role of maintaining intrinsic cell length control. Meanwhile, even after the development of a model that describes the MT dynami...
11 Pages(2750 words)Lab Report


Microtubules are polymers that have elongated tube-like monomer chains. These chains are helical like. α-βtubulin subunits make up the helical tube that has 13 protofilaments that are aligned in a parallel way. The subunits enable the tube-like monomers to link with each other using vertical and lateral bonds. The α-tubulin domain bind to the β-tubulin domain forming these bonds. Hence, the mechanical strength of the microtubules is enhanced. The Microtubule Organizing Centre mediates the nucleation of the microtubules. It comprises a ring complex of γ-TURC and γ-tubulin (Atkinson, 2014, 5870). The specific ring complex serve as the template of the 13 subunits of the microtubules. Microtubules grow from the minus to the positive s...
7 Pages(1750 words)Essay

The Dermal-Epidermal Junction

Some processes of excretion and gas exchange are also related to skin physiology (Marks, 1991; Powel, 1994). Skin also is the afferent part of the tactile analyzer (Pugliese, 2001). Nevertheless, the main function of the skin is protective. Different physiological mechanisms provide the protection from mechanical, thermal, chemical and biological factors (Pugliease, 2001). Thus the presence of melanocytes and skin pigmentation is an important factor in preventing UV caused lesions such as photodermatitis and skin cancer (Abdulla et al., 2005). The function of the cutaneous gland is important for normal microbiota persisting which is an antagonist to the potential dangerous transitory microorganisms. On the other hand, the skin cou...
32 Pages(8000 words)Dissertation

Hybridoma Technology

...Introduction A hybridoma, which can be considered as a hybrid cell, is produced by the injection of a specific antigen into a mouse, procuring the antibody-producing cell from the mouses spleen and the subsequent fusion of this cell with a cancerous immune cell called a myeloma cell. The hybrid cell, which is thus produced, can be cloned to produce many identical daughter clones. These daughter clones then secrete the immune cell product. Since these antibodies come from only one type of cell (the hybridoma cell) they are called monoclonal antibodies. The advantage of this process is that it can combine the qualities of the two different types of cells; the ability to grow continually, and to produce large amounts of pure... A hybridoma,...
6 Pages(1500 words)Coursework

Hormonal Secretion Peculiarities

... with specialized scaffold proteins. Scaffold proteins often interact also with membrane constituents, elements of the cytoskeleton, and adaptors mediating recruitment into clathrin-coated vesicles. They improve efficiency of signal transfer, facilitate interactions among different signal pathways, and control localization of signal proteins within a cell. Signal complexes are often associated with lipid raft domains of the plasma membrane. Scaffold proteins as well as signal proteins may be recruited from the cytosol to such membrane domains in part by insertion of lipid anchors, or by interaction of pleckstrin homology or other lipid-binding domains with head-groups of transiently formed phosphatidylinositol derivatives, , such as PIP2...
7 Pages(1750 words)Essay

Images of the Interior of the Body

...Introduction Ultrasound imaging, also known as ultrasound scanning or sonography is the viewing of images of the interior of the body by the use of high-frequency sound waves. Ultrasound scanning is useful to visualize various organs of the body as well as the vascular system. This essay briefly describes the uses of ultrasound in the imaging department and the vascular department. Uses of ultrasound in the imaging department 1. Head a. brain-in the newborn, because of the relatively thin parietal bones and patent fontanelles, one can use a conventional B-scanner or real-time sector scanner to get a cross sectional image of the body of the lateral and third ventricles. The thickness of the cerebral cortex can be measured... Ultrasound...
7 Pages(1750 words)Case Study

Oxaliplatin Genotoxicity on Human Lymphocytes

Oxaliplatin causes bulky DNA adducts. The absence of mismatch repair also indicated loss of p53 dependent apoptosis. CBMN could detect micronuclei which are indicative of DNA damage. Detection of micronuclei through CBMN indicated chromosome loss, breakage, rearrangement, nondisjunction, and apoptosis. All of these could mean specific genotoxic effects. These all are chromosomal aberrations, which may be caused by oxaliplatin. The mitotic catastrophe was a plausible mechanism of DNA toxicity caused by oxaliplatin through endocycling indicated in this experiment. This has been described as a process of mitotic failure induced by DNA damage leading to cell death. The other plausible mechanism that has been suggested is through decre...
15 Pages(3750 words)Research Paper

The Role of Histone Deacetylases in Human Cancer

The following essay summarizes the structure of chromatin with reference to the histones and discusses the post-translational modifications of these proteins.  The enzymes involved in acetylation/deacetylation of histones and the inhibitors of these enzymes are described in view of their applications in controlling human cancer.
The packaging unit of the chromatin is called nucleosome. The nucleosome is formed of a histone core of eight molecules of histones; two H2A, two H2B, two H3 and two H4. Each nucleosome consists of 1.75 superhelical turns (146 nucleotides length) of DNA wrapped around the core histone barrel-shaped octamers. The N-terminus of each octamer subunit is protruding free to be accessible for covalent...
11 Pages(2750 words)Coursework

The Human Cell

The researcher states that the chemical processes and complex interactions that occur inside the cell form the bridge between the nonliving and the living. Inside the human body, various processes that go on synchronized to provide efficient healthy functioning are interrelated and coupled for optimum functional output. This requires the organization at various levels. In complex living beings like humans, there are many levels of an organization, as opposed to single cellular organisms like bacteria. The study about the cell and its structure were revolutionized with the invention of the microscope. The discovery of microscope in 16th century encouraged scientists to study these basic building blocks. I...

20 Pages(5000 words)Term Paper
sponsored ads
We use cookies to create the best experience for you. Keep on browsing if you are OK with that, or find out how to manage cookies.

Let us find you another Essay on topic Immunolocalization of the microtubule cytoskeleton for FREE!

Contact Us