BioChem Lab final part 1

What is the stationary phase in SEC?

Mass of beads within the column

A spectrophotometer measures

A solutions absorbance at a specific wavelength

What did we use in the Protein Fractionation lab to separate proteins?

An acid and a salt

Why do set pipettes to maximum volume?

To preserve the internal spring

In the traditional method of pipetting, you collect the liquid while pressing at the:

First stop

0.777 ml is ____ uL

777.0

You can use P ___ to collect 0.2ml of solution

P2

___is the unit used to identify protein size

Daltons

In SEC, the biomolecules that are larger than the exclusion limit elutes out

Larger elutes first

In traditional method of pipetting, you collect the liquid while pressing to the ___ stop

First

___ is the type of chromatography where the column is filled with beads coated in "ligand"

Affinity

___ is the type of chromatography that is separation of proteins based on charge attraction and repulsion

Ionic exchange

Give one example of systematic error:

Pipettes are incorrectly calibrated

Give one example of a non-systematic error:

user errors such as wrong measurements

Typically ___ error is constant and can always be accounted for

Systematic

The ___ phase is usually the liquid phase that travels through the stationary phase pulling the compounds and separating them in column chromatography

Mobile

A protein has the lowest solubility at their ___ point

isoelectric

Salting our depends on the ___ of a protein

charge

Only ___ -toe shoes are acceptable to wear in the lab

closed

The ___ molecules move through the column first

Largest

Your labs are intended to expose you to concepts and methods of research and not to produce perfect results

True

Biuret assay is comprised of one redox reduction

True

When putting the pipette away, it should be set to its minimum setting

False

As soon as you collect the sample you are not supposed to allow the plunger to snap back in to position

True

After you plot your standard curve, you can only find the concentrations of unknown samples within the range of the curve

True

In affinity chromatography, it is important to use a buffer made of high concentration of salts or acid to finally elute out the protein of interest.

True

Running a blank is not necessary when you have to find the concentration of just one protein using a spectrophotometer

False

Forward pipetting is the best pipetting method to use when dealing with a viscous solution

True?

We keep proteins cold to prevent denaturing them

True

Non-systemic error is hard to account for as it is variable and usually caused by carelessness on the experimenter's part

True

SEC the liquid solution containing the protein sample, larger molecules elucidate first

mobile phase

SEC beads with pores trapping small proteins

stationary phase

The liquid is drawn while the plunger is at the second stop. This is to fully pull up the liquid and prevent the uptake of air

Reverse pipetting

A specific ligand is used as the stationary phase to bind to the desired protein

Affinity chromatography

Uses beads of the opposite charge of the desired protein

Ion exchange chromatography

Diagram of UV-spectrometer

Light source, entrance, dispersive element, wavelength, exit, sample, detector

If your pipette is giving inconsistent results, what could be the reason

Pushed to the second stop, side ways, incorrectly calibrated