Pregnancy Tests

Pregnancy Tests - wash proteins through column (small...

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Pregnancy Tests and Protein Biochemistry Western blot: run a SDS-PAGE gel or 2D gel, transfer the proteins to a “blot”, bock the unoccupied regions of the blot, add an antibody against your protein of interest (Ab 1 ), add a labeled antibody against the first antibody (Ab 2 ), image the blot; amplification by binding more than one secondary to each primary (also, cheaper) o Primary antibody: (Ab 1 ) against the protein of interest is washed across the pad; sticks only where there is the protein specific to that antibody o Secondary antibody: (Ab 2 ) against the first antibody is washed across the pad;sticks only to the primary antibody; can be wastefully coupled to a chromogenic enzyme or is made radioactive, making for easy identification; directed at a “constant domain” of the primary antibody, regardless of its specificity (can be mass-produced); allows for amplification of the signal; saves on reagents Chromatography: o Gel filtration chromatography: preparation by size; layer sample on column, add buffer to
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Unformatted text preview: wash proteins through column (small proteins attach to column), collect fractions (large proteins) o Ion exchange chromatography: preparation by charge; layer sample on column, negatively charged proteins attach to positively charged column, collect positively charged proteins, elute with NaCl, collect negatively charged proteins + Na+ o Affinity chromatography: preparation by specific binding; layer sample on column, add antibodies + pH 7 buffer, wash (collect protein which is not recognized by antibody), treat with pH 3 buffer, collect antibody-recognized protein Lateral flow immunochromatography: used in anthrax/pregnancy tests X-Ray crystallography: c rystalize your protein, blast it with a high-energy beam of x-rays, regular arrangement of electrons in the crystal diffracts the beam; use the diffraction pattern to determine an electron density map, thread the primary sequence through the electron density map...
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Pregnancy Tests - wash proteins through column (small...

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