Luminex xMAP AbC - Manual (Page 2)

xM
89-30
Micr
3
4
5
6
7
Acti
8
9
1
1
1
1
1
1
Incu
1
Micr
1
1
1
Cou
2
2
2
Incu
2
Micr
2
2
2
2
2
2
MAP
®
A
0000-00-361
rosphere W
3.  R
a. 1
b. 4
4.  D
5. WASH ST
a. 
b. 
c. 
d. 
6.  R
7.  R
ivation:
8.  A
9.  V
10.  V
11.  A
12.  A
13.  In
14.  A
15.  V
ubation #1:
16.  S
a. In
b. In
rosphere W
17. WASH ST
a. 
b. 
c. 
d. 
18.  R
19.  R
upling:
20.  A
21.  A
22.  V
ubation #2:
23.  S
a. R
b. R
rosphere W
24. WASH ST
a. 
b. 
c. 
d. 
25.  R
26.  R
27.  A
28.  V
29.  P
bC Kit
Rev. A
Wash #1:
Resuspend the
mL – vortex
mL – rotate f
Dispense the d
TEP – use on
 Place
 Remov
 Add 50
 Vortex
Repeat WASH
Remove liquid
Add Activation
Vortex and son
Vortex Sulfo-N
Add Sulfo-NH
Add 250 μL of
nvert EDC via
Add EDC solut
Vortex and son
hield from lig
ncubation star
ncubation end
Wash #2:
TEP – use on
 Place
 Remov
 Add 50
 Vortex
Repeat WASH
Remove liquid
Add Activation
Add Antibody
Vortex the reac
hield from lig
Rotation start t
Rotation end ti
Wash #3:
TEP – use on
 Place
 Remov
 Add 50
 Vortex
Repeat WASH
Remove liquid
Add 1mL of
W
Vortex and son
rotect from lig
Quick
e stock Bead
and sonicate
for 15 minutes
desired volum
e disposable
reaction tube
ve supernata
00 μL of Activ
x and sonicate
H STEP, for a
from Beads
n Buffer to re
nicate reactio
NHS tube for
S to reaction
Activation B
al and vortex f
tion to reactio
nicate reactio
ht and rotate
rt time:
d time:
e disposable
reaction tube
ve supernata
00 μL of Activ
x and sonicate
H STEP, for a
from Beads
n Buffer to th
y to the reactio
ction tube for
ht and rotate
time:
ime:
e disposable
reaction tube
ve supernata
00 μL of Was
x and sonicate
H STEP, for a
from Beads
ash Buffer to
nicate reactio
ght and store
Guide
s
for 10 secon
s at 15-30 rpm
me of Beads (
pipette per re
e into magneti
nt with transfe
vation Buffe
e reaction tub
total of two w
w/ magnetic
eaction tube (
n tube for 10
10 seconds
tube (from St
Buffer to 10 m
for 10-12 seco
on tube (from
n tube for 10
the reaction t
pipette per re
e into magneti
nt with transfe
vation Buffe
e reaction tub
total of three
w/ magnetic
he reaction tu
on tube (from
10 seconds
the reaction t
pipette per re
e into magneti
nt with transfe
sh Buffer into
e reaction tub
total of three
w/ magnetic
o the reaction
n tube for 10
at 2-8 ºC unt
ds
m
(from Step 2a
eaction tube
ic separator (
er pipette
r into reaction
be for 10 seco
washes
separator (1-
(from Step 2b
seconds
tep 2c )
mg vial of EDC
onds
Step 2d )
seconds
tube for 20 ±
eaction tube
ic separator (
er pipette
r into reaction
be for 10 seco
washes
separator (1-
be (from Step
Step 2e)
tube for 2 hou
eaction tube
ic separator (
er pipette
o reaction tube
be for 10 seco
washes
separator (1-
n tube
seconds
til needed
a ) to the react
1-2 min)
n tube
onds
2 min) and di
b )
C .
2 minutes @
1-2 min)
n tube
onds
2 min) and di
p 2f )
urs ± 5 minute
1-2 min)
e
onds
2 min) and di
tion tube
isposable pip
@ 15-30 rpm
isposable pip
es @ 15-30 r
isposable pip
ette
ette
rpm
ette
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