Sebia Hydragel

Question 1

intended use

Answer 1

separation of proteins in human serum and urine by electrophoresis

Question 2

6 major fractions

Answer 2

albumin
alpha 1
alpha 2
beta 1
beta 2
gamma

Question 3

Agarose gel

Answer 3

support medium for SPE

Question 4

When to discard agarose gel

Answer 4

crystal/precipitate forms, gel becomes soft (due to freezing the gel)

Bacterial or cold growth
Abnormal quantity of liquid present in gel box

Question 5

Buffered strips

Answer 5

Contains tris-barbital buffer (pH 8.5)

Function as EP buffer reservoir
ensure contact between gel and electrodes

Question 6

Discard buffered strips when

Answer 6

package is opened and the strips dry out

Question 7

Staining solution diluent

Answer 7

used for preparation of amidoblack staining solution

Question 8

Amidoblack stain

Answer 8

for staining gels after EP separation

Designed for only 10 stains (then change solution)

Question 9

Applicators

Answer 9

precept, single use

for sample application

Question 10

Filter papers

Answer 10

single use, thin absorbent paper pads for blotting excessive moisture off gel surface before sample application

Question 11

Reagents required (3)

Answer 11

destaining solution
Hydrasys wash solution
Fluidil

Question 12

Destaining solution

Answer 12

Removal of excess background stain from gels

Rinsing the staining compartment after wash step

Question 13

To neutralize acidity of destaining solution,

Answer 13

pour 15mL of 50%NaOH solution into the empty waste container

Question 14

Hydrasys wash solution

Answer 14

for cleaning hydrasys staining compartment

Discard working wash solution if it changes in appearance

Question 15

Fluidil

Answer 15

to dilute viscous or turbid samples (Eg. containing cryoglobulin or cryogel)

Must be free of precipitate

Question 16

Preferred sample for analysis

Answer 16

fresh serum

Fridge ASAP for up to 1 week

Question 17

What 2 things disappear after 3 days of storage of serum

Answer 17

beta 2, C3 complement

Question 18

What to add to frozen serum samples to improve storage stability

Answer 18

sodium azide 0.02 g/dL

Question 19

what to add to frozen urine samples to improve storage stability

Answer 19

HEPES 0.1M and sodium azide

Question 20

What may thawed samples present with

Answer 20

slight application marks due to protein and lipoprotein denaturation

older serum = greater shift

Question 21

why can’t you use a hemolyzed sample

Answer 21

increases alpha 2 and beta zones

Question 22

why should you avoid using plasma samples

Answer 22

fibrinogen gives band close to application point which might be seen as M peak

Offsets percentage of corresponding zones

Question 23

Concentrated urine

Answer 23

some have salt content which cause gel deformation during migration

Urine should be diluted

Question 24

Procedure

Answer 24

sample application
migration
drying
staining
destaining
final drying

Question 25

How much serum is put in each well

Answer 25

10 uL

Question 26

Temp during migration

Answer 26

65C for 10 min
50C (beep, lid unlocks)
cools to 20C when lid is opened

Question 27

4 things to check before stating

Answer 27

at least 300mL of staining solution
1 L of destaining solution
waste container empty
unused lines must be blocked