Test 2. Lab 8

Question 1

Describe rhodamine Phalloidin

Answer 1

Selectively stains F-actin

Excitiation/Emission: 540/565 nm

Superior to antibody staining

Optimal for fixed and permeabilized samples

Very widely cited fluorescent phalloidin conjugate

Question 2

*What does Rhodamine do in Rhodamine Phalloidin?

Answer 2

it is the color giving component that is attadched to phallotin that attaches to actin filmaments

Question 3

Define more about rhodamine phalloidin?

Answer 3

Two components – rhodamine and phalloidin

Rhodamine phalloidin is a high-affinity F-actin probe conjugated to the red-orange fluorescent dye, tetramethylrhodamine (TRITC).

Actin is the component of the cytoskeletal system that allows movement of cells and cellular processes.

Question 4

*compare and contrast rhodamine and phallodin?

Answer 4

RHODAMINE
Rhodamine is a family of related chemical compounds,fluoronedyes.

Rhodamine dyesare fluorescentand can thus be detected easily and inexpensively.

TRITC or Tetramethylrhodamine is a rhodamine derivative used in Rhodamine phalloidin.

PHALLODIN
Bicyclic peptide belonging to a family of toxins isolated from the deadlyAmanita phalloides”death cap” mushroom

Phallodin molecule binds to actin fibers

Fluorescently-labeled phalloidin has virtually identical binding properties with actin from different species including plants and animals.

Question 5

*____________ bind F - actin with high selectivity while (tetramethyl rhodamine) TRITC provides red orange fluorescence of unparallel brightnes and photo stability

Answer 5

phalloidin

Demonstrating very little nonspecific staining, Rhodamine phalloidin allows high-contrast

Question 6

Hoechst Fluorescent stain

Answer 6

Hoechst stainsare part of a family of bluefluorescentdyesused tostainDNA.

Bis - benzimidesoriginally developed by theHoechst AG, a German company.

The dyesHoechst 33258and Hoechst 33342are the ones most commonly used and they have similarexcitation/emission spectra.

Question 7

*Hoechst Fluorescent Stain- binds preferentially with what regions of DNA

Answer 7

AT regions

Question 8

*What is the wavelengths of Rhodamine Phalloidin?

Answer 8

540/565

Question 9

What type of actin does rhodamine phalloidin stain?

Answer 9

f actin

Question 10

• What wavelengths are used for Hoistein dye?

Answer 10

33258/ 33342