Protein Microarrays Protein Microarrays
Visit Molecular Station for a great article on Antibody and Protein Microarrays Posjetite molekularnu stanica za velik članak na Antitijelo i protein Microarrays
Protein Chips: Protein Čipovi:
Protein Microarrays can be based on any ligand-binding assay that relies on the formation of product with an immobilized capture molecule and a target molecule present in the solution. Microarrays protein može biti zasnovana na bilo kojoj ligand-binding esej koji se oslanja na stvaranje proizvoda s immobilized capture molekula i ciljne molekule prisutna u rješenje. These assays can be miniturized and placed on a protein microarray chip. Ovi eseji može biti miniturized i na protein microarray čip.
Protein microarrays are now becoming very popular due to their possible future applications in the study of nucleic acid-protein, protein-protein, ligand-receptor, drug-protein target, and enzyme-substrate interactions. Protein microarrays su sada postaje vrlo popularan zbog svoje moguće buduće primjene u istraživanju nucleic acid-protein, protein-protein, ligand-receptor, droga-protein target, i interakcije enzim-podloga.
Types of Capture Molecules for Protein Microarray Chips. Vrste Capture molekula za protein Microarray Chips. To analyze protein interactions with other molecules, protein microarrays can have various types of molecules immobilized on the slide surface to act as capture molecules in the protein microarray assay. Za analizu protein interakcije s drugim molekulama, protein microarrays mogu imati različite vrste molekula immobilized na slajd površine djelovati kao hvatanje molekula u protein microarray esej.
Figure 1 Illustrates the most common protein array, the antibody microarray. Slika 1 pokazuje najčešće protein array, antitijela microarray. Antibodies are spotted onto a glass slide. Protutijela su uočena na staklu slajd. Two cell lysates with proteins labeled with Cy3 and Cy5 (for example, cancer vs normal) are added as a solution onto the protein array. Dva cell lysates proteini s etiketom s Cy3 i Cy5 (na primjer, rak vs normal) su dodano kao rješenje na protein array. Signals are detected from the two different signals. Signali su otkrivena je iz dva različita signala. Data is computed allowing an analysis of how much dye-labeled protein the antibodies on each spot are binding. Data je kompjutorska omogućava analizu koliko pigment-protein s etiketom protutijela na svakom mjestu su obvezujući. This information allows one to determine whether the protein expression is up-regulated in cancer, down-regulated or unchanged. Ova informacija omogućuje da utvrdi da li se protein expression je up-regulirani u raka, down-regulirani ili nepromijenjene.
Figure 2. Slika 2. a) Demonstrates protein arrays which are based on microarray analysis of antigen-antibody interactions. a) nizovi Demonstrates protein koji se temelje na microarray analysis of antigen-antitijelo interakcije. Antigens are spotted onto glass slides. Antigeni su uočena na staklu dijapozitiva. Antibodies which are tagged bind to antigens and emit fluorescent signal (shown as the yellow star) which can then be detected from the spot on the array. Protutijela kojima su označene povezano s antigeni i uputiti fluorescencija signal (kao što je prikazano u žute zvjezdice) koji se zatim biti otkrivena od točke na polje.
b) Shows a protein microarray composed of spots which act as sandwich immunoassays. b) Pokazuje microarray protein sastavljen od mjesta koje djeluju kao sendvič immunoassays. Antibodies are spotted onto the chips and cell lysate or a solution is applied to the slide. Protutijela su uočena na čips i cell lysate ili rješenje je primijenjeno u slajd. Antibodies are incubated with the solution, with later washing to remove non-specific binding. Protutijela su incubated s rješenjem, s kasnije perilica za uklanjanje ne-posebna obvezujuća. The sandwich complexes formed emit a detectable signal from the spot. U sendvič naselja formirana uputiti jednom detektabilni signal iz točke.
c) A protein-protein interaction protein microarray where proteins are spotted onto a protein chip, and labeled proteins are added to the chip and incubated to determine where they may bind and interact. c) A protein-protein interakcije proteina protein microarray gdje su uočena na protein čip, i etiketom proteini su dodane na čip i incubated kako bi utvrdio gdje oni svibanj svežemo i interakciju. A detectable signal is emitted from binding spots. A detektabilni signal se emitiraju iz obvezujuća mjesta.
d) Nucleic Acid-protein interaction array, (here a DNA-protein interaction chip). d) nucleic acid-protein interakcije polje, (ovdje DNA-protein interakcije čip). Nucleic acids such as DNA are spotted onto a glass slide. Nucleic Acids, kao što su DNA su uočena na staklu slajd. This could be transcriptional binding sites for example. Proteins such as fluorescently labeled DNA-binding transcription factors in solution are added to the chip and incubated on the array. To može biti obvezujuća transcriptional stranice za primjer. Proteini, kao što su fluorescently etiketom DNA-binding transkripcijski činitelji u rješenje su dodane na čip i incubated na polje. A detectable signal is emitted from the DNA spots where the proteins bind. A detektabilni signal se emitiraju iz DNA mjesta gdje se proteini svezati.
e) If you have a set of ligands and you want to find the receptors the bind to, you can use a receptor-ligand protein chip as displayed. Receptor proteins are spotted on glass slides. e) Ako imate niz liganada i želite pronaći receptori u svežu, možete koristiti jedan receptor protein-ligand čip kao prikazani. receptor proteini su uočena na staklu dijapozitiva. Fluorescently labeled ligands are added to the chip. Fluorescently označenom kao liganada su dodane na čip. Binding of ligand to receptors causes the emission of a detectable signal which pinpoints the interacting spots (ie which protein did the ligand bind to). Obvezujuća za ligand do receptora uzrokuje zračenje od detektabilni signal koji se međusobno pinpoints mjesta (tj. protein koji je ligand povezano s).
f) To test for enzyme-subrates specificity one can employ an enzyme-substrate protein microarray. f) Da bi test za enzim-subrates specifičnost može se zaposliti jedan enzim-podloga protein microarray. This chip is composed of descrete nano-wells in which enzymes are bound. Ovaj čip se sastoji od nano-descrete bunara u koji su dužni enzimi. Substrate is added into the nano-wells and signal emission is detected from the nano-wells in order to assay for enzyme function. Podloga je dodano u nano-bunara i signal emisije je otkrivena od nano-bunare kako bi se esej za enzim funkcija.
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