Tag: Protein

Introduction

Protein, DNA, and RNA can all be separated from one another using an external electric field and this laboratory technique. As the gel or matrix employed includes a given size and enables just a particular molecule, the separation of molecules occurs on the basis of charge and size. Only very small particles, such as atoms or ions, may fit through the pores. By doing so, molecular separation is possible.

When electricity flows, ions can migrate to various poles. In electrophoresis, the movement of anions is known as anaphoresis and that of cations is known as cataphoresis. When a current is applied, proteins migrate to the positive poles, where they may be more easily separated.

What is Electrophoresis?

Electrophoresis is the process by which a current drives charged molecules to separate poles. Molecules with a positive charge will go away from the cathode and towards the anode. Negatively charged molecules will seek out a cathode, which is the opposite pole of an electrode.

As proteins and nucleic acids are negatively charged, they gravitate towards the cathode. Because to its association with electricity and ion motion, electro kinetics is another name for this field. Capillary electrophoresis and slab electrophoresis are the two most common forms. Gel and paper electrophoresis are examples of capillary electrophoresis, while Iso-electro focusing and Zone electrophoresis are examples of slab electrophoresis.

Gel Electrophoresis

Applications of Electrophoresis

Electrophoresis is applied in many fields because of its feasibility and inexpensive mode. Some of its applications are,

DNA Fragmentation and DNA Analysis 

Gel electrophoresis is the process used for the separation of DNA fragments based on their size. The approximate size of DNA is also obtained in this process

Protein Detection

Gel electrophoresis is again employed for the detection of proteins. Proteins have higher migratory power and that is employed for its detection. By the application of external current, the charged particles of proteins will migrate towards their opposite electrodes. As protein is negatively charged it will migrate towards the cathode. It can also use the liquid matrix for the separation. The majority of protein applications can be achieved by the use of this process. They include purity determination and purification of proteins.

Testing of Antibodies

Immune electrophoresis detects antibodies. Antibodies on a gel do it. blood test. If the sample contains the antigen, an antigen-antibody complex will form. IgA, IgM, etc. are easily diagnosed.

Factors affecting the Process of Electrophoresis

Some factors can substantially affect the process of electrophoresis. They are,

Electric Field: Electric field is responsible for the movement of charged particles towards the oppositely charged electrodes. So it plays an important role in the process of electrophoresis.

 Sample features/details: The features of the sample such as size, shape, and charge have an important role since it is a size-based and charge-based separation. If the charge of a molecule is high the rate of migration will also high. But if the size increases the rate will decrease.

Buffer: For stabilizing the pH of the medium buffer solution is necessary the use of zwitterionic buffers will be much better.

Supporting Medium: It plays a predominant role since the medium can alter the rate of migration by having some adsorption property. Electro osmosis will also result due to some mediums. So selection of a proper support medium will increase the rate of migration.

 How does gel electrophoresis work?

Gel electrophoresis is a type of electrophoresis in which charged particles or ions of biological molecules can be separated by the application of an external field. They move through a gel hence the name gel electrophoresis is used. And its movement is called migration. And the migration of ions is to oppositely charged electrodes. The electric current for this process is applied across the gel. The gel acts as a sieve since it contains small pores that can only allow small-sized molecules. DNA fragments are distinguished using this electrophoresis. And DNA is a negatively charged species so it will migrate to the rds cathode.

What is agarose gel electrophoresis?

One of agar’s main components is a matrix for the electric current-based synthesis of macromolecules. Agarose gel is commonly employed as a matrix in the separation of DNA and big protein molecules because it is simple to cast, has a good pore size, and has a good gel strength. It has several charged groups and aids in water movement in the opposite direction of DNA movement towards the anode. A larger concentration of agarose gel is required for the separation of tiny molecules.

What are the advantages of electrophoresis?

The advantages of electrophoresis are,

• The mode of operation of this technique is easy.
• Short time is only needed.
• Needs only a low sample.
• The consumption of electrolytes during the process is low.
• Can be used as an analytical tool and also as a synthesis tool.
• The bulk amount of proteins shall be separated.
• Inexpensive apparatus.

Summary

Samples are separated by size and charge using electrophoresis. It is medium dependent, and the various forms it takes are defined by the medium used. Several types of electrophoresis in tubes and capillaries. Gel electrophoresis, zone electrophoresis, etc., are also included. Gel electrophoresis stands out because it is used to separate proteins, DNA, RNA, and other biomolecules. Gels aid the separation process since only molecules of a certain size may pass through them; if an electric field is applied, the molecules in question will align themselves into a distinct band, making them easy to identify. The supercoiled structure of agarose gel makes it a viable alternative matrix. Electrophoresis has various benefits due to being a cheap and efficient method.

 Frequently Asked Questions

1. What is the difference between SDS-PAGE and Native PAGE?

Ans. SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) is a technique used to separate proteins based on their size and charge. Native PAGE (polyacrylamide gel electrophoresis) is a technique used to separate proteins based on their size and shape. 

2. What is Capillary Electrophoretic Enzyme Assay?

Ans. Applications of the analytical separative technique known as capillary electrophoresis (CE) extend far beyond the realm of enzymatic research.

In an open capillary, the substrate and the product are separated by size and charge in an electric field.

3. What is the haemoglobin electrophoresis test?

Ans: It is an electrophoresis test that can be used to the identification of the amount of haemoglobin present in the sample blood.

Introduction

A system of ductless glands that secrete hormones is known as an endocrine gland system. In the blood, hormones are carried directly to their target organs as chemical messengers. As hormones are synthesized in certain tissues, they trigger a cascade of events that allow for certain cellular responses. Adrenal glands are located above the kidney. They are divided into regions cortex and medulla. Both regions produce hormones that regulate electrolyte balance.

What are Adrenal Glands?

The body contains two adrenal glands. These are located above each kidney. They are a part of our endocrine system. Despite their small size, the adrenal glands are quite important for your body’s hormone-related functions. As a result, elements that affect your adrenal glands may have a big impact on your general health. If you believe you might have an adrenal problem, see a doctor.

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Location of Gland

The adrenal glands are endocrine glands that are located over the lateral surface of each kidney’s upper poles. The adrenal glands are located at the back of the abdomen, between the kidney and the ribs. They are triangular, and the parietal peritoneum only covers the front of them. The right gland is pyramid-shaped, while the left gland is moderately-lunar.

                                    

Hormones Secreted by Adrenal Gland 

• The gland’s outer layer is called the cortex. It generates both cortisol and aldosterone. The medulla is the gland’s most interior area. It generates both adrenaline and noradrenaline.
• Epinephrine, also referred to as adrenaline, is a hormone that responds swiftly to anxiety by quickening the heartbeat and raising blood sugar levels in the body. 
• Corticosterone controls the immune system and reduces inflammation in conjunction with hydrocortisone. 
• Cortisol: With the help of cortisol, the body controls and utilizes fats, protein, and carbohydrates. In addition, it weakens the immune system and affects physiology, which the body uses to help it cope with worry. 
• Aldosterone: helps the kidneys keep the blood and tissues of the body’s healthy salt chloride level. 
• Norepinephrine, also referred to as noradrenaline, works in conjunction with epinephrine to respond to anxiety. Its primary function is to prepare the body and mind for combat.

Function

• The adrenal glands are responsible for producing several hormones that help control blood pressure, produce several reproductive hormones, and maintain the proper level of salt in our blood and cells. 
• The adrenal glands’ release of stress-inducing hormones causes one of the most well-known reactions, the “conflict of the panic response.” 
• When and how rapidly an adolescent develops their reproductive system, helping during birth 
• Our endocrine system’s pituitary gland, also known as the “master gland,” controls our adrenal glands. The main controller of your endocrine glands is the pituitary gland, which is located inside your skull. Abnormal signals may interfere with the number of hormones your pituitary gland advises your adrenal glands to generate.

Disease-related to the Adrenal Gland

Adrenal gland dysfunction might manifest when 

• When your master gland is unable to efficiently regulate your hormone production, the adrenal glands may form benign or non-malignant tumours or harmful or cancerous tumours. 
• Can suffer from disorders of the adrenal glands. 
• People are born with particular gene diseases.

Cushing’s Disease

People with Cushing’s syndrome have excessive cortisol production from their adrenal glands. The most common cause of this is prolonged use of high dosages of corticosteroids. Lupus, arthritis, and pneumonia are just a few of the disorders that are treated with corticosteroids. They have cortisol-like effects throughout the body. 

Symptoms:

• The round face, the fat accumulation at the base of the neck, and the fat bulge between the arms.
• Skinny arms and thighs
• Gaining weight unnecessarily
• Prone to scarring skin
• On the hips, chest, and stomach, there are wide, violet stretch marks.

Addison Disease

A rare autoimmune ailment called Addison’s disease may develop when your adrenal glands don’t produce enough cortisol or aldosterone. This self-harm sickness may cause your immune cells to attack the tissues of your adrenal glands. 

Symptoms: 

• Arterial pressure is high despite the weight 
• Loss or gain without cause. 
• Hyperglycemia or intolerance to glucose 
• low amounts of potassium 
• Discomfort, stress, or panic episodes

Congenital Adrenal Hyperplasia (CAH)

Extremely little cortisol is synthesized in congenital adrenal hyperplasia (CAH), a genetic disease. Furthermore, persons who have this condition may also have additional hormonal issues in which their bodies create too much testosterone but not enough aldosterone.

Symptoms: 

• Early acne, body hair, beards, or loud voices (women)
• Rapid growth in children
• Sterility due to underarm hair

Adrenocortical Carcinoma

Adrenocortical carcinoma is a malignant tumour that often develops on the exterior of the adrenal gland. Oftentimes, this type of tumour is only found after it has spread to the body’s other systems over generations.

Summary

The adrenal glands are located above the kidney. Despite their small size, the adrenal glands are quite important for your body’s hormone-related functions. The gland’s outer layer is called the cortex. It generates both cortisol and aldosterone. The adrenal glands’ release of stress-inducing hormones causes one of the most well-known reactions, the “conflict of the panic response.” When your master gland is unable to efficiently regulate your hormone production, the adrenal glands may form benign or non-malignant tumours or harmful or cancerous tumours.

Frequently Asked Questions

1. Is it necessary to have both Adrenal Glands to Survive?
Ans. Since the adrenal glands are necessary for human survival, removing both of them (which is quite unusual) forces the patient to take medications and hormone replacements.

2. Is it Possible to live with just one Adrenal Gland?
Ans. Your doctor will administer painkillers to you. Your body can function regularly if your adrenal gland is working properly. If both of your adrenal glands were lost or if your remaining adrenal gland isn’t functioning properly, you might take medication every day to restore the hormones they were making.

3. Do the Adrenal Glands play a role in Digestion?
Ans. The adrenal glands release the stress hormone cortisone to inhibit bodily processes like the immune response and digestion that are not necessary for immediate survival.

4. What are Hormones that make us Drained?
Ans. The adrenal glands also create the hormone aldosterone, which is problematic since it helps regulate your body’s normal fluid and electrolyte balances. As adrenal depletion increases, it makes your body generate less aldosterone, which results in dryness and electrolyte imbalances.