Biochem cram

What is the principle of Differential centrifugation

Differential centrifugation separates cell components based on differences in their size and density by spinning them at different speeds.

What does differential centrifugation determine? What is the use?

It determines the size and density of cell components.

What is the advantage of differential centrifugation?

It is a quick and simple way to separate cell components.

What is the limitation of differential centrifugation?

It does not completely separate cell components, so the fractions may still contain a mixture of different organelles

What is the principle of Dialysis

Dialysis separates molecules based on their size using a semipermeable membrane. Small molecules pass through the membrane, while larger molecules are kept inside.

what does dialysis determine/what is the use?

remove small molecules (like salts) from a solution while keeping larger molecules, such as proteins.

what is the advantage of dialysis?

It can remove small molecules without damaging or losing the larger molecules, like proteins.

what is the limitation of dialysis?

It is slow and cannot separate molecules that are similar in size.

what is the principle of salting out?

Salting out separates proteins based on their solubility in high salt concentrations.

what does salting out determine/what is the use?

It is used to separate and concentrate proteins based on their different solubilities in salt.

what is the advantage of salting out?

It is simple, inexpensive, and can separate large amounts of proteins.

what is the limitation of salting out?

It does not produce very pure proteins, so the proteins may still be mixed together.

what is the principle of ultracentrifugation?

separates molecules or cell components based on their size, shape, and density using very high-speed spinning.

what does ultracentrifugation determine/what is the use?

It is used to separate and study molecules or cell components based on their size, shape, and density.

what is the advantage of ultracentrifugation?

It provides better separation of molecules and cell components than regular centrifugation.

what is the limitation of ultracentrifugation?

It is expensive, requires specialized equipment, and can take a long time to perform.

what is the principle of gel filtration?

separates molecules based on size. Larger molecules move through the column faster, while smaller molecules get trapped in the gel and move more slowly

what does gel filtration determine/what is the use

It is used to separate proteins and estimate their size.

what is the limit of gel filtration?

It has a limited ability to separate molecules that are very similar in size.

what is the advantage of gel filtration?

It is gentle and does not usually damage proteins, and it can also estimate protein size.

what is the principle Of ion exchange chromatography

separates proteins based on their charge. Proteins bind to the column based on their charge and are then released by changing the salt concentration or pH.

what does ionic exchange chromatography determine/what is the use?

It is used to separate and purify proteins based on their electrical charge.

what is the advantage of ion exchange chromatography?

It can separate proteins very precisely based on their charge and can produce highly purified proteins.

what is the limitation of ion exchange chromatography?

It only works well when proteins have different charges, and changes in pH or salt concentration can affect the proteins.

what is the principle of affinity chromatography?

Affinity chromatography separates proteins based on their specific binding to a molecule attached to the column.

what does affinity chromatography determine/what is the use?

It is used to isolate and purify a specific protein from a mixture based on its unique binding ability.

what is the advantage of affinity chromatography?

It is highly specific and can produce a very pure protein in one step.

what is the limitation of affinity chromatography?

It can be expensive, and it requires a specific binding molecule for the protein you want to isolate.

what is the principle of HPLC?

HPLC separates molecules based on how strongly they interact with the column as they are pushed through it by high pressure.

what does HPLC determine/what is the use?

It is used to separate, identify, and measure the amount of different molecules in a mixture.

what is the advantage of HPLC?

It is fast, highly accurate, and can separate very similar molecules.

what is the limitation of HPLC?

It is expensive, requires specialized equipment, and can be difficult to operate.

what is the principle of isoelectric focusing

separates proteins based on their isoelectric point (pI), the pH at which a protein has no net charge.

What does isoelectric focusing determine/what is the use?

It is used to separate and identify proteins based on their isoelectric point (pI).

what Is the advantage of isoelectric focusing?

It can separate proteins with very similar charges very precisely.

what is the limitation of isoelectric focusing?

It can be difficult to perform and does not work well for very small or very large proteins.

what is the principle of 2-D page Gel Electrophoresis?

separates proteins in two steps: first by their charge (isoelectric point) and then by their size.

what does two dimensional gel electrophoresis determine/what is the use

It is used to separate and compare many proteins in a sample based on both their charge and size.

what is the advantage of two dimensional gel electrophoresis

It provides very detailed separation of proteins because it separates them by both charge and size.

what is the limitation of two dimensional gel electrophoresis

It is time-consuming, technically difficult, and some proteins may not separate well.

what is the principle of SDS page?

separates proteins based on size. SDS gives proteins a similar negative charge, so smaller proteins move faster through the gel than larger proteins.

what does SDS Page determine/what is the use?

It is used to separate proteins based on their size and determine their approximate molecular weight.

what is the advantage of SDS page?

It separates proteins by size very effectively and can estimate their molecular weight.

what is the limitation of SDS page?

It can denature proteins, so you cannot study their normal shape or function.

what is the principle of western blotting?

separates proteins by size using SDS-PAGE and then uses antibodies to specifically detect a particular protein.

what does western blading determine/what is the use?

used to identify and measure the amount of a specific protein in a sample using antibodies.

what is the advantage of western blotting?

It is highly specific and can detect a particular protein even when it is mixed with many other proteins.

what is the limitation of western blotting?

It can be time-consuming and requires specific antibodies for the protein you want to detect.

what is the principle of co-immunoprecipitation?

uses an antibody to pull down a specific protein along with other proteins that are interacting with it.

what does co-immunoprecipitation determine/what is the use?

It is used to determine whether two or more proteins interact with each other in a cell.

what is the advantage of co-immunoprecipitation?

It can identify protein-protein interactions in a cell under relatively natural conditions.

what is a limitation of immunoprecipitation?

that it requires a specific antibody that binds well to the target protein.

what is the principle of ELISA

the specific binding between an antigen and an antibody, followed by an enzyme-driven color change to detect and measure target molecules

what does ELISA determine/what is the use?

uses antibodies linked to an enzyme to detect a specific protein or molecule. The enzyme produces a color change, which indicates how much of the target is present.

what is the limitation of ELISA

A limitation is that it requires specific antibodies and can produce false-positive or false-negative results.

what is the principle of mass spectrometry

separates molecules based on their mass-to-charge ratio (m/z) and measures them to identify the molecules in a sample.

what does mass spectrometry Determined/what is the use?

It is used to identify molecules and determine their molecular mass.

what are the advantages of mass spectrometry

It is highly sensitive and accurate, and it can identify and measure very small amounts of molecules.

what are the limitations of mass spectrometry

It is expensive, requires specialized equipment, and can be complex to operate and interpret.

what is the principle of NMR spectroscopy

uses a magnetic field and radio waves to measure the behavior of atomic nuclei, which produces signals that can be used to study the structure of molecules.

what does NMR spectroscopy determine

determines the structure of molecules by showing the chemical environment of their atoms. It can be used to identify compounds and determine how atoms are connected.

what is an advantage of NMR spectroscopy?

It can determine the structure of molecules without destroying the sample.

what is limitation of NMR spectroscopy?

It requires expensive, specialized equipment and relatively large amounts of sample.

what is the principle of cryo-EM

uses an electron microscope to image samples that have been rapidly frozen, allowing molecules to be studied in a near-natural state.

what does cryo-EM determine/what is the use?

It is used to determine the 3D structure and shape of large molecules, such as proteins and protein complexes.

what are the advantages of cryo-EM

It can determine the 3D structure of large proteins and complexes without needing to crystallize them, and it preserves them in a near-natural state.

what are the limitations of cryo-EM

It is expensive, requires specialized equipment, and can be difficult to prepare and analyze samples.

what is the principle of x-ray crystallography

uses X-rays to create a diffraction pattern from a crystallized molecule, which is used to determine its 3D structure.

what does x-ray crystallography determine/what is the use?

It is used to determine the 3D structure of molecules, especially proteins, at a very detailed level.

what are the advantages of x-ray crystallography?

It can provide very high-resolution, detailed 3D structures of molecules, showing the precise positions of atoms.

what are the limitations of x-ray crystallography?

It requires the molecule to form a good crystal, which can be difficult, and it can be time-consuming to prepare the sample.

what is the advantage of ELISA

It is highly sensitive and specific, and it can detect and measure small amounts of a target molecule.