Lecture 4 SDS PAGE and Western

During the protein analysis lab, the purpose of SDS PAGE is to separate proteins by __________.

size

The equation used for calculating the concentration of unknown samples in the lab is y = 1.6722x. Here, 'y' represents ____________.

absorance

The __________ gel is where proteins are first introduced into the gel before entering the resolving gel.

stacking

The main purpose of adding sodium dodecyl sulfate (SDS) during SDS PAGE is to provide a ____________ charge to the proteins.

net negative

The standard protein used for comparison in SDS PAGE is called ____________.

Kaleidoscope

The technique used to transfer proteins from a gel to a membrane is called ____________.

blotting

For Western blotting, the primary antibodies bind specifically to the ____________ of interest.

antigen

In monoclonal antibody production, the B cells are fused with ____________ cells to create hybridomas.

cancer

The secondary antibodies used in Western blotting are created against the _____________ region of the primary antibody.

constant

A ____________ antibody recognizes multiple epitopes on an antigen, providing greater variability but reduced specificity

polyclonal

When using enzymatic labels, the substrate reacts with the enzyme to produce a ____________ that can be detected visually.

precipitate

To avoid nonspecific binding of antibodies in Western blotting, a blocking protein like ____________ can be used.

milk

After running the gel, it is important to check for the ____________ pattern of proteins to ensure sufficient loading during SDS PAGE.

banding

Antibodies are proteins produced by ____________ cells in response to antigens.

B

what does SDS PAGE stand for

sodium dodecyl sulfate polyacrylamide gel electrophoresis

what does BSA stand for

bovine serum albumin

it is essential to handle ___ with care due to its neurotoxic properties, emphasizing safety precautions in the lab environment

acrylamide

composition of gel, forming a porous matrix for protein seperation

acrylamide, bis-acrylamide polymerized under the influence of ammonium persulfate and TEMED

___ are critical in determining the pore sizes suitable for protein sizes, the smaller the proteins the higher this is

gel percentages

These offer high specificity by binding to a single epitope and are produced from hybridoma cells, which are derived from spleen cells fused with myeloma cells.

monoclonal antibodies

___ can be labeled directly or indirectly for signal detection

antigens

____ are often utilized for signal amplification and flexibility

secondary antibodies