Lab 6- Polyacrylamide Gel Electrophoresis (SDS PAGE) Procedure

Polyacrylamide Gel

Mesh-like matrix for protein separation.

Disulfide Bonds

Covalent bonds disrupted by SDS and reducing agents.

Sample Buffer

Buffer mixed with protein samples for loading.

Running Buffer

Buffer facilitating electrophoresis in the gel.

SDS/PAGE

Electrophoresis method for protein separation by mass.

Sodium Dodecyl Sulfate (SDS)

Anionic detergent used to denature proteins.

Polypeptide Chain Length

Length of protein chains affecting migration distance.

Relative Migration Distance (Rf)

Distance traveled by proteins inversely related to MW.

Molecular Weight (MW)

Mass of proteins influencing their movement in gel.

Protein Standards

Known MW proteins used for estimating sample MW.

Centrifuge

Device used to separate components by density.

Electrophoresis Power Supply

Provides constant voltage for protein migration.

Coomassie Staining

Method for visualizing proteins on gel.

Heating Block

Equipment used to denature protein samples.

Microcentrifuge Tubes

Small tubes for sample preparation and storage.

Protein Concentration

Amount of protein needed for visualization on gel.

Anode

Positive electrode where proteins migrate during electrophoresis.

Cathode

Negative electrode in the electrophoresis setup.

Electrophoresis Chamber

Container holding gel and buffers during electrophoresis.

Staining Detection Method

Technique used to visualize proteins after electrophoresis.

Voltage Setting

Constant voltage of 150 volts for electrophoresis.

Sample Loading Volume

Volume of protein sample loaded into gel lanes.

Heating Temperature

Samples heated to 95°C for denaturation.

Protein Lysates

Cell extracts containing proteins for analysis.