Jv Uhcl Biochem Lab Test 1 - Focused

Front 1

--- Equations ---
1) Making pH buffers
2) X-solutions
3) Centrifugation
4) Extinction Coefficient

Back 1

1) Henderson–Hasselbalch equation
2) V1C1 = V2C2
3) Stokes equation
4) Beer-Lambert Law

Front 2

pH = pKa + [***/***]

Back 2

[A- / HA]
[base / acid]

Front 3

10 = [*** / ***]
Significance?

Back 3

[10 / 1]
!!! 11 Units !!!

Front 4

100mL of 10% NaCl(w/v)

Back 4

10g NaCl
add water up to 100mL

Front 5

100mL of 10% H3O(v/v)

Back 5

10mL H3O
90mL of water

Front 6

Percent Solution Types...

Back 6

w/v
v/v

Front 7

pI?
pKa?

Back 7

isoelectric point
acid dissociation constant

Front 8

--- Different Protein Elution Volumes ---
1) Due to??

Back 8

Different size/weight

Front 9

Stokes equation: larger particles.....

Back 9

sediment faster

Front 10

1) Vmax Equation?
2) Vmax Units?
3) Vmax AKA?
4) Km Equation?

Back 10

1) 1 / Y intercept
2) μmol/min
3) Total Activity
4) -1 / X intercept

Front 11

Total Activity AKA ***

Back 11

Vmax

Front 12

--- Specific Activity ---
Equation?

Back 12

Total Activity(Vmax)
------------------
Total Protein

Front 13

--- Specific Activity Analysis ---
It's assumed "kinetic analysis" and "enzyme concentration" tests share the same ***.

Back 13

[E] per mL

Front 14

--- Finding Enzyme Concentration ---
1) *** range matched w/std concentrations.
2) Compensate for sample ***.

Back 14

1) linear
2) dilution

Front 15

--- Finding Enzyme Concentration ---
- 0.01mL Enzyme matched w/ 2.5μg/mL STD -
1) Concentration of Enzyme per mL?

Back 15

1) 250μg/mL

Front 16

--- DNA/RNA Absorbance ---
1) Major peak at ***
2) Minor peak at ***

Back 16

1) 260 nm
2) 280 nm

Front 17

--- Protein Absorbance ---
1) Major peak at ***
2) Minor peak at ***

Back 17

1) 280 nm
2) 260 nm

Front 18

--- Protein Concentration Quantification ---
1) Method used in Question 4B?
2) Benefit of this method?
3) Why direct 280nm absorbance not used?

Back 18

1) Bradford dye-binding procedure
2) Highly sensitive/specific for protein
3) DNA/RNA contamination

Front 19

--- Density & Centrifugation ---
1) Two methods mentioned?
2) Methods differ in their *** ***.
3) However, both use *** or ***.

Back 19

1) Step / Linear Gradient
2) Density Gradients
3) Percoll or sucrose

Front 20

--- Step Gradient Banding ---
1) Bands form at step t***
2) Where at in this question?
3) Yes, ***.

Back 20

1) transitions
2) Very top, between 2 layers, very bottom
3) Cell pellet @ bottom.

Front 21

--- Linear Gradient Banding ---
1) Bands form at c*** ***...
2) No ***.

Back 21

1) corresponding densities
2) Cell pellet @ bottom.

Front 22

--- Density & Centrifugation ---
1) Particles must be...... to their...

Back 22

1) Centrifuged to their equilibrium densities

Front 23

--- Purification Table ---
Purification Fold?

Back 23

Fn Specific Activity
---------------------------
F1 Specific Activity

Front 24

--- Purification Table ---
% Yield?

Back 24

Fn Total Activity(Vmax)
-----------------------
F1 Total Activity(Vmax)

Front 25

--- Purification Table ---
1) Don't forget to consider *** and ***.

Back 25

1) dilution --- units

Front 26

--- Beer-Lambert Equation ---
1) Formula?
2) Variables? Units?

Back 26

A = εcl
A = absorbance
ε = Extinction Coefficient
c = concentration (M)
l = path length of cuvette (cm)